卵黄高磷蛋白的分离纯化、结构表征及功能特性研究
摘要
卵黄高磷蛋白是鸡蛋蛋黄中主要的磷蛋白,独特的氨基酸组成和高度磷酸化使其具有很多优良的物化和生物学特性,生物信息学分析推测卵黄高磷蛋白可能具有调控生物矿化的功能。本课题首先分离制备高纯度的卵黄高磷蛋白,对其结构进行表征,鉴定磷酸化位点并结合生物信息学分析,挖掘其潜在功能,最后研究卵黄高磷蛋白调控矿化的机制和活性位点。本课题为卵黄高磷蛋白在仿生矿化中的应用奠定基础。主要研究内容和结果如下：
(1)建立一种条件温和、高效简单的分离卵黄高磷蛋白的新方法,将鸡蛋蛋黄依次经等质量蒸馏水和0.17mol/LNaCl稀释离心除去蛋黄浆质,获得蛋黄颗粒,用10%1.74mol/LNaCl溶解后,添加3%w/w PEG6000并调整溶液pH至4.0,离心后透析冻干,制备的卵黄高磷蛋白(Pvti)电泳鉴定纯度为99%,得率为47%。采用该方法分别从海兰褐和白莱航鸡蛋中分离制备卵黄高磷蛋白,HPLC测定纯度分别达93.14%和91.80%,得率分别为63.93%和58.89%。进一步分析表明,这两个品种中的卵黄高磷蛋白的含量、组成成分和二级构象均无明显差异,且与Sigma的一致。此外,使用离子交换层析法制备不含多价金属离子的卵黄高磷蛋白,最佳条件为：Pvti经Na2EDTA处理后,采用梯度增加NaCl浓度(0、0.35、0.5mol/L)的Tris-HCl缓冲液(0.05mol/L, pH7.5)在DEAE柱中洗脱,收集洗脱峰经透析冻干,得到纯化的不含多价金属离子的卵黄高磷蛋白(PVtf)。
(2)开展卵黄高磷蛋白结构的系统分析,研究显示卵黄高磷蛋白的氨基酸组成含有高达30%的丝氨酸(Ser),富含酸性氨基酸(Asp. Glu)和碱性氨基酸(Arg、His),含有极少量的芳香族氨基酸(Tyr、Cys、 Trp)。使用红外光谱(FTIR)和圆二色谱(CD)分析卵黄高磷蛋白二级结构主要为无规卷曲和p-折叠结构,不含金属离子的卵黄高磷蛋白p-转角结构增多。LC-ESI-MS/MS (LTQ)生物质谱技术结合生物信息学预测分析表明卵黄高磷蛋白含有87个磷酸化位点(83个P-Ser和4个P-Thr)。利用生物信息学分析9个物种来源的11种卵黄高磷蛋白以及来源于人的5种磷酸化蛋白质,进行氨基酸组成、多序列比对和进化树分析,显示均具有高丝氨酸含量,且富含酸性氨基酸的特点。鸡卵黄高磷蛋白与牙本质磷蛋白(DPP)的结构及特性相似,进化关系较近,意味着卵黄高磷蛋白可能具有与牙本质磷蛋白相似的生物功能,即在生物矿化过程中发挥重要的调控作用。
(3)利用钙离子浓度计(ISE)、等温滴定量热仪(ITC)、CD和荧光光谱方法,分别从化学、热力学以及结构信息方面,较为全面地研究了卵黄高磷蛋白与钙离子的相互作用。在中性和弱碱性条件下,卵黄高磷蛋白与钙离子相互作用存在高亲和性和低亲和性两类结合位点。高亲和性位点结合常数约为104mol-1,每摩尔卵黄高磷蛋白约结合30摩尔的钙离子,该结合为焓驱动反应,卵黄高磷蛋白的构象变化为无序和p-转角结构增加,而p-折叠结构减少,主要作用力为静电、氢键或范德华力；而低亲和性位点结合常数和结合位点数并不恒定,有大量的钙离子累积在饱和钙的分子或聚集区域的附近,该结合为熵驱动反应,卵黄高磷蛋白的p-折叠结构增加,主要作用力为疏水作用力；而在酸性条件下,卵黄高磷蛋白与钙离子相互作用为熵驱动的吸热反应,主要作用力为弱的疏水相互作用。
(4)研究仿生矿化体系中卵黄高磷蛋白对磷酸钙矿物晶相转化的影响。FTIR、X射线衍射(XRD)、扫描电镜(SEM)和能谱分析(EDS)表征矿物的结构、组成以及表面微观形貌,结果表明在该体系中生成的磷酸钙矿物通过“溶解再结晶”途径从磷酸氢钙(DCPD)转变为羟基磷灰石(HAP)。卵黄高磷蛋白可以极大地促进晶相转化,使转化过程由6h缩短至0.5h以内。磷酸钙矿物的FTIR和XRD图谱显示出蛋白质特征峰,且矿化后滤液中蛋白含量降至原来的0.5%,表明卵黄高磷蛋白参与了磷酸钙矿物的形成,成为矿物的组成部分之一。不同添加顺序的实验结果显示,卵黄高磷蛋白在DCPD形成之前添加才能够发挥其促进作用,表明与游离钙离子的互作是其调控磷酸钙矿化的关键。作为对照的BSA亦对晶相转化具有促进作用,但其作用较弱且参与模式不同,BSA参与磷酸钙矿物晶相转化后期阶段,而卵黄高磷蛋白参与晶相转化起始阶段,卵黄高磷蛋白在磷酸钙矿化过程中的奇特作用与其高度的磷酸化密切相关。
(5)使用0.1、0.2、0.3、0.4mol/L NaOH溶液处理卵黄高磷蛋白(Pv),经超滤分离得到不同去磷酸化的卵黄高磷蛋白：T1、T2、T3和T4,去磷酸化程度分别为2.98%、19.46%、43.39%和71.07%。并采用pH-stat体系研究了其对矿化的影响。卵黄高磷蛋白促进矿化的效果呈浓度剂量关系。通过CD、 FTIR、 XRD、 SEM和荧光光谱研究不同去磷酸化程度的卵黄高磷蛋白对矿化的影响,结果表明磷酸化在矿化反应中起到很重要的作用。去磷酸化后的卵黄高磷蛋白p-折叠结构增加,可以提供矿化模板。去磷酸化程度低的卵黄高磷蛋白由于无序结构减少,吸附矿物能力减弱,含磷量少,结合钙离子能力减弱,从而促进矿化效果减弱。但高浓度碱液处理卵黄高磷蛋白使其水解为了卵黄高磷蛋白磷酸肽,由于活性区域暴露,成核位点增加,显著的促进矿化。总之,促进矿化效果强弱顺序依次为T4>T3>卵黄高磷蛋白﹥T2﹥T1﹥Control。
(6)通过碱液部分去磷酸化后再用胰蛋白酶酶解卵黄高磷蛋白,经DEAE离子交换色谱和Sepharose G-25凝胶过滤色谱分离纯化卵黄高磷蛋白磷酸肽(PPP)各组分,SDS-PAGE和CD分析显示PPPO和PPP1可能为卵黄高磷蛋白酶解的小分子片段(N端或C端),而PPP3和PPP4二级结构相似,只是PPP4折叠更多,且PPP3多含10kDa的小分子肽。利用pH-stat体系研究卵黄高磷蛋白磷酸肽各组分对矿化效果的影响,通过CD、 FTIR、 XRD、 SEM和荧光光谱研究发现PPPO、 PPP1、 PPP4促进效果并没有卵黄高磷蛋白强,原因为PPP0和PPP1是小分子肽,不参与矿化；同时,PPP4折叠太多,结构太紧凑,使得促进矿化能力减弱。而PPP3暴露了活性区域,因此促进效果最显著。促进矿化反应效果强弱顺序依次为：PPP3>卵黄高磷蛋白﹥PPP4﹥PPP1﹥Control﹥PPPO。经LTQ MS/MS鉴定可知促进矿化的活性区域为卵黄高磷蛋白核心区域中的D1165-R1258。
Phosvitin is the major phosphoprotein from hen egg yolk. Due to its unique amino acid composition and highly phosphorylation, phosvitin possess many excellent physical and biological properties. And phosvitin was speculated to possess regulation properties of biological mineralization through the bioinformatics analysis. In our research, highly purified phosvitin was firstly prepared, then characterized its structure, identified the phosphorylation sites, and excavated the potential function with combing the bioinformatics analysis, lastly investigated the regulation mechanism of phosvitin in the mineralization and clarified its active sites. This study will lay a foundation for the application of phosvitin in biomimetic mineralization. The main research contents and results are as follows:
An attempt was made to develop a new protocol for preparing phosvitin with high efficiency and simplicity in a mild condition. Hen egg yolk sequentially diluted with equal mass of distilled water and0.17M NaCl, to obtain yolk granules after removing the yolk plasma by centrifugation. Granules was dissolved with10%1.74M NaCl, added3%w/w PEG6000and the solution was adjusted to pH4.0, desalted by dialysis after centrifugation and lyophilized. The purity of phosvitin (Pvti) was99%, which identified by SDS-PAGE, and yield was47%. Based on this protocol, phosvitin was prepared from egg yolk of Hy-Line Brown and White Leghorn, the purity examined by HPLC was93.14%and91.80%, respectively; and the yield was63.93%and58.89%, respectively. Further study showed that there had hardly significant between Hy-Line Brown and White Leghorn in the content, composition and secondary structure of phosvitin, they were all consistent with Sigma. Furthermore, Polyvalent metal-free phosvitin was purified through ion-exchange chromatography, the optimal condition was as follows: Pvti treated with Na2EDTA was eluted with an increasing gradient of NaCl (0,0.35and0.5mol/L) in Tris-HCl buffer (0.05mol/L, pH7.5) by DEAE column, the eluate was collected, polyvalent metal-free phosvitin (Pvtf) was obtained through dialysis against distilled water and lyophilized.
(2) Systematic analysis the structure of phosvitin, the studies showed that amino acid composition of phosvitin contain up to30%Ser, rich in acidic amino acids (Asp, Glu) and basic amino acids (Arg, His), and contain a very small amount of the aromatic amino acids (Tyr, Cys and Trp). The conformation of phosvitin was mainly random coil and (3-sheet structure while metal-free phosvitin increased the β-turn content, by using Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD). Phosvitin possessed87phosphorylation sites (83P-Ser and4P-Thr), which were identified by LC-ESI-MS/MS (LTQ) and combined the prediction results by bioinformatics analysis. In addition, bioinformatics analyzed the11types of phosvitin from9different species and5types of phosphorylated protein from human through amino acid compositions, multiple sequence alignment and phylogenetic analysis, the results revealed that they all rich in acidic amino acids. Phosvitin and dentin phosphophoryn (DPP) have similar structure and properties, meanwhile the evolutionary relationships was close, this suggested phosvitin may have similar biological functional activity with DPP, can play an important regulation role in the process of bio mineralization.
(3) Comprehensive study of the interaction of phosvitin with calcium ions from the chemical, thermodynamic and structural aspects, by using calcium ion selective electrode (ISE), isothermal titration calorimetry (ITC), CD and fluorescence spectroscopy, respectively. The results showed that, under neutral and alkaline conditions, there existed two classes of binding sites in the interaction between phosvitin and calcium: high affinity site and weak affinity site. The binding constant of high affinity is about104mol-1, and binding sites were nearly30mol of calcium per mole of phosvitin. This reaction was driven by enthalpy, the conformation of phosvitin increased random and (3-sheet structure, while decreased β-turn structure, the main interaction force was electrostatic, hydrogen bonds or Van der Waals. Meanwhile, the binding constant and the binding site of low affinity were not constant, since a large amount of calcium ions accumulated in the vicinity of the calcium saturation region or the molecule aggregation, the binding reaction was driven by entropy, the conformation of phosvitin increased β-sheet structure, and main interaction force was hydrophobic force. However, under acidic condition, the interaction between phosvitin and calcium was entropy-driven endothermic reaction, and the main interaction force was weak hydrophobic force.
(4) Effect of phosvitin on the phase transformation of calcium phosphate mineral was investigated in the biomimetic system. The mineral structure, composition and surface morphology was characterized by FTIR, X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS). The results showed that the calcium phosphate mineral transformed from dicalcium phosphate dihydrate (DCPD) to hydroxyapatite (HAP), through the "dissolution-recrystallization" pathway in this system. Phosvitin significantly accelerated the phase transformation, since the transform process was shortened from6h to less than0.5h. Besides, The characteristic peaks of protein was occurred in the FTIR spectra and the XRD patterns of calcium phosphate mineral, and the phosvitin concentration in the filtrate during the mineralization was reduced to0.5%, indicated that phosvitin was involved in the phase transformation of calcium phosphate mineral and incorporated into the mineral. The results of the addition sequence of substrates on the phase transformation of calcium phosphate showed that, Phosvitin played its role only if adding phosvitin before DCPD formed, indicates that the interaction between phosvitin and the free calcium ions was the key to promote its regulation mineralization of calcium phosphate. BSA used as a control, also promoted the phase transformation. However, compared with phosvitin, its effect was weak and was involved in different modes. BSA involved in the later phase transformation stages of calcium phosphate mineral while phosvitin participate the initial phase transformation. The peculiar role of phosvitin in the calcium phosphate mineralization process was closely related to its degree of phosphorylation.
(5) Different dephosphorylation degrees of phosvitin were accomplished by treating phosvitin (Pv) with0.1,0.2,0.3,0.4mol/L NaOH solution. T1, T2, T3and T4were obtained through ultrafiltration, the dephosphorylation degree was2.98%,19.46%,43.39%,71.07%, respectively. Their effect on the mineralization were studied by using a pH-stat system. Phosvitin promote mineralization depended on concentration-dose relationship. Effect of different dephosphorylation degrees of phosvitin on the mineralization was studied by CD, FTIR, XRD, SEM, fluorescence spectroscopy. The results showed that phosphorylation was very important in the mineralization reaction. In addition, the dephosphorylation phosvitin increased its P-sheet structure, providing the template of mineralization. Due to its random structure reduced, low dephosphorylated levels of phosvitin decreased its adsorption capacity on mineral, decreased the calcium binding capacity as less phosphorus, and thereby weakened the effect of promotion on the mineralization. However, phosvitin could hydrolyzed to phosvitin peptides by high concentration of alkaline solution, the active regions were exposed, thus increased the nucleation sites, and significantly promoted mineralization. In summary, efficiency of promoting mineralization order was T4﹥T3﹥Pv﹥T2﹥T1﹥Control.
(6) Phosvitin was partially dephosphorylated by alkaline and then digested with trypsin. The phosvitin phosphopeptides (PPP) fractions were separated and purified through DEAE ion-exchange chromatography and Sepharose G-25gel filtration chromatography. SDS-PAGE and CD analysis showed that PPPO and PPP1were possible the small molecule fragments (N-or C-terminal) after enzymatic hydrolysis, while PPP3and PPP4had similar secondary structure, but PPP4had more compact and β-sheet structure, while PPP3contained10kDa peptides. Effect of phosvitin phosphopeptides fractions on the mineralization by means of FTIR, XRD, SEM, fluorescence spectroscopy in pH-stat system. The results showed that, as compared with Pv, PPPO, PPP1and PPP4decreased its promoting efficiency, because PPPO and PPP1were small peptides, and not involved in the mineralization; meanwhile, PPP4owed too much β-sheet structure resulted in more compact, thus weakened the promotion effect. PPP3exposed its active region, thus the promoted effect was the most significant. The mineralization reaction rate was as follows: PPP3﹥Pv﹥PPP4﹥PPP1﹥Control﹥PPPO. The active region of promotion mineralization was D1165-R1258in the core region of phosvitin by LTQ MS/MS identification.
(1)建立一种条件温和、高效简单的分离卵黄高磷蛋白的新方法,将鸡蛋蛋黄依次经等质量蒸馏水和0.17mol/LNaCl稀释离心除去蛋黄浆质,获得蛋黄颗粒,用10%1.74mol/LNaCl溶解后,添加3%w/w PEG6000并调整溶液pH至4.0,离心后透析冻干,制备的卵黄高磷蛋白(Pvti)电泳鉴定纯度为99%,得率为47%。采用该方法分别从海兰褐和白莱航鸡蛋中分离制备卵黄高磷蛋白,HPLC测定纯度分别达93.14%和91.80%,得率分别为63.93%和58.89%。进一步分析表明,这两个品种中的卵黄高磷蛋白的含量、组成成分和二级构象均无明显差异,且与Sigma的一致。此外,使用离子交换层析法制备不含多价金属离子的卵黄高磷蛋白,最佳条件为：Pvti经Na2EDTA处理后,采用梯度增加NaCl浓度(0、0.35、0.5mol/L)的Tris-HCl缓冲液(0.05mol/L, pH7.5)在DEAE柱中洗脱,收集洗脱峰经透析冻干,得到纯化的不含多价金属离子的卵黄高磷蛋白(PVtf)。
(2)开展卵黄高磷蛋白结构的系统分析,研究显示卵黄高磷蛋白的氨基酸组成含有高达30%的丝氨酸(Ser),富含酸性氨基酸(Asp. Glu)和碱性氨基酸(Arg、His),含有极少量的芳香族氨基酸(Tyr、Cys、 Trp)。使用红外光谱(FTIR)和圆二色谱(CD)分析卵黄高磷蛋白二级结构主要为无规卷曲和p-折叠结构,不含金属离子的卵黄高磷蛋白p-转角结构增多。LC-ESI-MS/MS (LTQ)生物质谱技术结合生物信息学预测分析表明卵黄高磷蛋白含有87个磷酸化位点(83个P-Ser和4个P-Thr)。利用生物信息学分析9个物种来源的11种卵黄高磷蛋白以及来源于人的5种磷酸化蛋白质,进行氨基酸组成、多序列比对和进化树分析,显示均具有高丝氨酸含量,且富含酸性氨基酸的特点。鸡卵黄高磷蛋白与牙本质磷蛋白(DPP)的结构及特性相似,进化关系较近,意味着卵黄高磷蛋白可能具有与牙本质磷蛋白相似的生物功能,即在生物矿化过程中发挥重要的调控作用。
(3)利用钙离子浓度计(ISE)、等温滴定量热仪(ITC)、CD和荧光光谱方法,分别从化学、热力学以及结构信息方面,较为全面地研究了卵黄高磷蛋白与钙离子的相互作用。在中性和弱碱性条件下,卵黄高磷蛋白与钙离子相互作用存在高亲和性和低亲和性两类结合位点。高亲和性位点结合常数约为104mol-1,每摩尔卵黄高磷蛋白约结合30摩尔的钙离子,该结合为焓驱动反应,卵黄高磷蛋白的构象变化为无序和p-转角结构增加,而p-折叠结构减少,主要作用力为静电、氢键或范德华力；而低亲和性位点结合常数和结合位点数并不恒定,有大量的钙离子累积在饱和钙的分子或聚集区域的附近,该结合为熵驱动反应,卵黄高磷蛋白的p-折叠结构增加,主要作用力为疏水作用力；而在酸性条件下,卵黄高磷蛋白与钙离子相互作用为熵驱动的吸热反应,主要作用力为弱的疏水相互作用。
(4)研究仿生矿化体系中卵黄高磷蛋白对磷酸钙矿物晶相转化的影响。FTIR、X射线衍射(XRD)、扫描电镜(SEM)和能谱分析(EDS)表征矿物的结构、组成以及表面微观形貌,结果表明在该体系中生成的磷酸钙矿物通过“溶解再结晶”途径从磷酸氢钙(DCPD)转变为羟基磷灰石(HAP)。卵黄高磷蛋白可以极大地促进晶相转化,使转化过程由6h缩短至0.5h以内。磷酸钙矿物的FTIR和XRD图谱显示出蛋白质特征峰,且矿化后滤液中蛋白含量降至原来的0.5%,表明卵黄高磷蛋白参与了磷酸钙矿物的形成,成为矿物的组成部分之一。不同添加顺序的实验结果显示,卵黄高磷蛋白在DCPD形成之前添加才能够发挥其促进作用,表明与游离钙离子的互作是其调控磷酸钙矿化的关键。作为对照的BSA亦对晶相转化具有促进作用,但其作用较弱且参与模式不同,BSA参与磷酸钙矿物晶相转化后期阶段,而卵黄高磷蛋白参与晶相转化起始阶段,卵黄高磷蛋白在磷酸钙矿化过程中的奇特作用与其高度的磷酸化密切相关。
(5)使用0.1、0.2、0.3、0.4mol/L NaOH溶液处理卵黄高磷蛋白(Pv),经超滤分离得到不同去磷酸化的卵黄高磷蛋白：T1、T2、T3和T4,去磷酸化程度分别为2.98%、19.46%、43.39%和71.07%。并采用pH-stat体系研究了其对矿化的影响。卵黄高磷蛋白促进矿化的效果呈浓度剂量关系。通过CD、 FTIR、 XRD、 SEM和荧光光谱研究不同去磷酸化程度的卵黄高磷蛋白对矿化的影响,结果表明磷酸化在矿化反应中起到很重要的作用。去磷酸化后的卵黄高磷蛋白p-折叠结构增加,可以提供矿化模板。去磷酸化程度低的卵黄高磷蛋白由于无序结构减少,吸附矿物能力减弱,含磷量少,结合钙离子能力减弱,从而促进矿化效果减弱。但高浓度碱液处理卵黄高磷蛋白使其水解为了卵黄高磷蛋白磷酸肽,由于活性区域暴露,成核位点增加,显著的促进矿化。总之,促进矿化效果强弱顺序依次为T4>T3>卵黄高磷蛋白﹥T2﹥T1﹥Control。
(6)通过碱液部分去磷酸化后再用胰蛋白酶酶解卵黄高磷蛋白,经DEAE离子交换色谱和Sepharose G-25凝胶过滤色谱分离纯化卵黄高磷蛋白磷酸肽(PPP)各组分,SDS-PAGE和CD分析显示PPPO和PPP1可能为卵黄高磷蛋白酶解的小分子片段(N端或C端),而PPP3和PPP4二级结构相似,只是PPP4折叠更多,且PPP3多含10kDa的小分子肽。利用pH-stat体系研究卵黄高磷蛋白磷酸肽各组分对矿化效果的影响,通过CD、 FTIR、 XRD、 SEM和荧光光谱研究发现PPPO、 PPP1、 PPP4促进效果并没有卵黄高磷蛋白强,原因为PPP0和PPP1是小分子肽,不参与矿化；同时,PPP4折叠太多,结构太紧凑,使得促进矿化能力减弱。而PPP3暴露了活性区域,因此促进效果最显著。促进矿化反应效果强弱顺序依次为：PPP3>卵黄高磷蛋白﹥PPP4﹥PPP1﹥Control﹥PPPO。经LTQ MS/MS鉴定可知促进矿化的活性区域为卵黄高磷蛋白核心区域中的D1165-R1258。
Phosvitin is the major phosphoprotein from hen egg yolk. Due to its unique amino acid composition and highly phosphorylation, phosvitin possess many excellent physical and biological properties. And phosvitin was speculated to possess regulation properties of biological mineralization through the bioinformatics analysis. In our research, highly purified phosvitin was firstly prepared, then characterized its structure, identified the phosphorylation sites, and excavated the potential function with combing the bioinformatics analysis, lastly investigated the regulation mechanism of phosvitin in the mineralization and clarified its active sites. This study will lay a foundation for the application of phosvitin in biomimetic mineralization. The main research contents and results are as follows:
An attempt was made to develop a new protocol for preparing phosvitin with high efficiency and simplicity in a mild condition. Hen egg yolk sequentially diluted with equal mass of distilled water and0.17M NaCl, to obtain yolk granules after removing the yolk plasma by centrifugation. Granules was dissolved with10%1.74M NaCl, added3%w/w PEG6000and the solution was adjusted to pH4.0, desalted by dialysis after centrifugation and lyophilized. The purity of phosvitin (Pvti) was99%, which identified by SDS-PAGE, and yield was47%. Based on this protocol, phosvitin was prepared from egg yolk of Hy-Line Brown and White Leghorn, the purity examined by HPLC was93.14%and91.80%, respectively; and the yield was63.93%and58.89%, respectively. Further study showed that there had hardly significant between Hy-Line Brown and White Leghorn in the content, composition and secondary structure of phosvitin, they were all consistent with Sigma. Furthermore, Polyvalent metal-free phosvitin was purified through ion-exchange chromatography, the optimal condition was as follows: Pvti treated with Na2EDTA was eluted with an increasing gradient of NaCl (0,0.35and0.5mol/L) in Tris-HCl buffer (0.05mol/L, pH7.5) by DEAE column, the eluate was collected, polyvalent metal-free phosvitin (Pvtf) was obtained through dialysis against distilled water and lyophilized.
(2) Systematic analysis the structure of phosvitin, the studies showed that amino acid composition of phosvitin contain up to30%Ser, rich in acidic amino acids (Asp, Glu) and basic amino acids (Arg, His), and contain a very small amount of the aromatic amino acids (Tyr, Cys and Trp). The conformation of phosvitin was mainly random coil and (3-sheet structure while metal-free phosvitin increased the β-turn content, by using Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD). Phosvitin possessed87phosphorylation sites (83P-Ser and4P-Thr), which were identified by LC-ESI-MS/MS (LTQ) and combined the prediction results by bioinformatics analysis. In addition, bioinformatics analyzed the11types of phosvitin from9different species and5types of phosphorylated protein from human through amino acid compositions, multiple sequence alignment and phylogenetic analysis, the results revealed that they all rich in acidic amino acids. Phosvitin and dentin phosphophoryn (DPP) have similar structure and properties, meanwhile the evolutionary relationships was close, this suggested phosvitin may have similar biological functional activity with DPP, can play an important regulation role in the process of bio mineralization.
(3) Comprehensive study of the interaction of phosvitin with calcium ions from the chemical, thermodynamic and structural aspects, by using calcium ion selective electrode (ISE), isothermal titration calorimetry (ITC), CD and fluorescence spectroscopy, respectively. The results showed that, under neutral and alkaline conditions, there existed two classes of binding sites in the interaction between phosvitin and calcium: high affinity site and weak affinity site. The binding constant of high affinity is about104mol-1, and binding sites were nearly30mol of calcium per mole of phosvitin. This reaction was driven by enthalpy, the conformation of phosvitin increased random and (3-sheet structure, while decreased β-turn structure, the main interaction force was electrostatic, hydrogen bonds or Van der Waals. Meanwhile, the binding constant and the binding site of low affinity were not constant, since a large amount of calcium ions accumulated in the vicinity of the calcium saturation region or the molecule aggregation, the binding reaction was driven by entropy, the conformation of phosvitin increased β-sheet structure, and main interaction force was hydrophobic force. However, under acidic condition, the interaction between phosvitin and calcium was entropy-driven endothermic reaction, and the main interaction force was weak hydrophobic force.
(4) Effect of phosvitin on the phase transformation of calcium phosphate mineral was investigated in the biomimetic system. The mineral structure, composition and surface morphology was characterized by FTIR, X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS). The results showed that the calcium phosphate mineral transformed from dicalcium phosphate dihydrate (DCPD) to hydroxyapatite (HAP), through the "dissolution-recrystallization" pathway in this system. Phosvitin significantly accelerated the phase transformation, since the transform process was shortened from6h to less than0.5h. Besides, The characteristic peaks of protein was occurred in the FTIR spectra and the XRD patterns of calcium phosphate mineral, and the phosvitin concentration in the filtrate during the mineralization was reduced to0.5%, indicated that phosvitin was involved in the phase transformation of calcium phosphate mineral and incorporated into the mineral. The results of the addition sequence of substrates on the phase transformation of calcium phosphate showed that, Phosvitin played its role only if adding phosvitin before DCPD formed, indicates that the interaction between phosvitin and the free calcium ions was the key to promote its regulation mineralization of calcium phosphate. BSA used as a control, also promoted the phase transformation. However, compared with phosvitin, its effect was weak and was involved in different modes. BSA involved in the later phase transformation stages of calcium phosphate mineral while phosvitin participate the initial phase transformation. The peculiar role of phosvitin in the calcium phosphate mineralization process was closely related to its degree of phosphorylation.
(5) Different dephosphorylation degrees of phosvitin were accomplished by treating phosvitin (Pv) with0.1,0.2,0.3,0.4mol/L NaOH solution. T1, T2, T3and T4were obtained through ultrafiltration, the dephosphorylation degree was2.98%,19.46%,43.39%,71.07%, respectively. Their effect on the mineralization were studied by using a pH-stat system. Phosvitin promote mineralization depended on concentration-dose relationship. Effect of different dephosphorylation degrees of phosvitin on the mineralization was studied by CD, FTIR, XRD, SEM, fluorescence spectroscopy. The results showed that phosphorylation was very important in the mineralization reaction. In addition, the dephosphorylation phosvitin increased its P-sheet structure, providing the template of mineralization. Due to its random structure reduced, low dephosphorylated levels of phosvitin decreased its adsorption capacity on mineral, decreased the calcium binding capacity as less phosphorus, and thereby weakened the effect of promotion on the mineralization. However, phosvitin could hydrolyzed to phosvitin peptides by high concentration of alkaline solution, the active regions were exposed, thus increased the nucleation sites, and significantly promoted mineralization. In summary, efficiency of promoting mineralization order was T4﹥T3﹥Pv﹥T2﹥T1﹥Control.
(6) Phosvitin was partially dephosphorylated by alkaline and then digested with trypsin. The phosvitin phosphopeptides (PPP) fractions were separated and purified through DEAE ion-exchange chromatography and Sepharose G-25gel filtration chromatography. SDS-PAGE and CD analysis showed that PPPO and PPP1were possible the small molecule fragments (N-or C-terminal) after enzymatic hydrolysis, while PPP3and PPP4had similar secondary structure, but PPP4had more compact and β-sheet structure, while PPP3contained10kDa peptides. Effect of phosvitin phosphopeptides fractions on the mineralization by means of FTIR, XRD, SEM, fluorescence spectroscopy in pH-stat system. The results showed that, as compared with Pv, PPPO, PPP1and PPP4decreased its promoting efficiency, because PPPO and PPP1were small peptides, and not involved in the mineralization; meanwhile, PPP4owed too much β-sheet structure resulted in more compact, thus weakened the promotion effect. PPP3exposed its active region, thus the promoted effect was the most significant. The mineralization reaction rate was as follows: PPP3﹥Pv﹥PPP4﹥PPP1﹥Control﹥PPPO. The active region of promotion mineralization was D1165-R1258in the core region of phosvitin by LTQ MS/MS identification.
引文
1.马杰.鸡卵黄高磷蛋白中和内毒素作用的研究[硕士学位论文].青岛：中国海洋大学图书馆,2012
2.倪莉,王璋,许时婴.鸡蛋中卵黄高磷蛋白的提取.无锡轻工大学学报，1999：57-61
3.徐彩娜.卵黄高磷蛋白磷酸肽的抗炎机理研究[博士学位论文].长春：吉林大学,2012
4. Abdelkebir K, Morin-Grognet S, Gaudiere F, Coquerel G, Labat B, Atmani H, Ladam G. Biomimetic layer-by-layer templates for calcium phosphate biomineralization. Acta Biomater, 2012,8:3419-3428
5. Abe Y, Itoh T, Adachi S. Fractionation and characterization of hen's egg yolk phosvitin. J Food Sci, 1982, 47: 1903-1907
6. Albright K, Gordon D, Cotterill O. Release of iron from phosvitin by heat and food additives. J Food Sci,1984,49:78-81
7. Allerton SE, Perlmann GE. Chemical characterization of the phosphoprotein phosvitin. JBiol Chem, 1965, 240: 3892-3898
8. Anton M. Composition and structure of hen egg yolk. In: ed. Bioactive egg compounds:Springer Berlin Heidelberg, 2007:1-6
9. Anton M, Castellani O, Guerin-Dubiard C. Phosvitin. In: ed. Bioactive Egg Compounds:Springer Berlin Heidelberg, 2007:17-24
10. Arifuzzaman S, Rohani S. Experimental study of brushite precipitation. J Cryst Growth,2004,267:624-634
11. Atha DH, Ingham KC. Mechanism of precipitation of proteins by polyethylene glycols. Analysis in terms of excluded volume. JBiol Chem, 1981,256:12108
12. Baht G, O'Young J, Borovina A, Chen H, Tye C, Karttunen M, Lajoie G, Hunter G, Goldberg H. Phosphorylation of Ser136 is critical for potent bone sialoprotein-mediated nucleation of hydroxyapatite crystals. Biochem J, 2010, 428: 385-395
13. Bassani G, Farruggia B, Nerli B, Romanini D, Pico G. Porcine pancreatic lipase partition in potassium phosphate-polyethylene glycol aqueous two-phase systems. J Chromatogr B, 2007, 859:222-228
14. Bee L, Cotterill O. Ion-exchange chromatography and electrophoresis of egg yolk proteins. J Food Sci, 1979,44:656-667
15. Belhomme C, David-Briand E, Guerin-Dubiard C, Vie V, Anton M. Phosvitin-calcium aggregation and organization at the air-water interface. Colloids Surf, B, 2008,63:12-20
16. Belhomme C, David-Briand E, Ropers M-H, Guerin-Dubiard C, Anton M. Interfacial characteristics of spread films of hen egg yolk phosvitin at the air-water interface: Interrelation with its charge and aggregation state. Food Hydrocolloids,2007,21:896-905
17. Bos MA, Vliet Tv. Interfacial rheological properties of adsorbed protein layers and surfactants: a review. Adv Colloid Interface Sci, 2001,91:437-471
18. Boskey AL. Mineral-matrix interactions in bone and cartilage. Clin Orthop Relat R,1992,281:244-274
19. Boskey AL. Matrix proteins and mineralization: an overview. Connect Tissue Res, 1996,35:357-363
20. Brockbank RL, Vogel HJ. Structure of the oligosaccharide of hen phosvitin as determined by two-dimensional 1H NMR of the intact glycoprotein. Biochemistry, 1990,29:5574-5583
21. Burke E, Guo Y, Colon L, Rahima M, Veis A, Nancollas G. Influence of polyaspartic acid and phosphophoryn on octacalcium phosphate growth kinetics. Colloids Surf, B, 2000, 17:49-57
22. Burley RW, Cook WH. Isolation and composition of avian egg yolk granules and their constituent α-and β-lipovitellins. Can J Biochem Physiol, 1961,39: 1295-1307
23. Byrne BM, Gruber M, Ab G. The evolution of egg yolk proteins. Prog Biophys Mol Bio,1989,53:33-69
24. Byrne BM, Van het Schip AD, Van de Klundert JA, Arnberg AC, Gruber M, Ab G. Amino acid sequence of phosvitin derived from the nucleotide sequence of part of the chicken vitellogenin gene. Biochemistry, 1984, 23:4275-4279
25. Carbonaro M, Nucara A. Secondary structure of food proteins by Fourier transform spectroscopy in the mid-infrared region. Amino Acids, 2010, 38: 679-690
26. Carnevali O, Cionna C, Tosti L, Lubzens E, Maradonna F. Role of cathepsins in ovarian follicle growth and maturation. Gen Comp Endocr, 2006, 146:195-203
27. Castellani O, Belhomme C, David-Briand E, Guerin-Dubiard C, Anton M. Oil-in-water emulsion properties and interfacial characteristics of hen egg yolk phosvitin. Food Hydrocolloids, 2006, 20:35-43
28. Castellani O, Belhomme C, David-Briand E, Guerin-Dubiard C, Anton M. The role of metal ions in emulsion characteristics and flocculation behaviour of phosvitin-stabilised emulsions. Food Hydrocolloids, 2008, 22:1243-1253
29. Castellani O, David-Briand E, Guerin-Dubiard C, Anton M. Effect of aggregation and sodium salt on emulsifying properties of egg yolk phosvitin. Food Hydrocolloids,2005,19:769-776
30. Castellani O, Guerin-Dubiard C, David-Briand E, Anton M. Influence of physicochemical conditions and technological treatments on the iron binding capacity of egg yolk phosvitin. Food Chem, 2004,85:569-577
31. Castellani O, Martinet V, David-Briand E, Guerin-Dubiard C, Anton M. Egg yolk phosvitin: Preparation of metal-free purified protein by fast protein liquid chromatography using aqueous solvents. J Chromatogr B, 2003,791:273-284
32. Causeret D, Matringe E, Lorient D. Ionic strength and pH effects on composition and microstructure of yolk granules. JFood Sci, 1991,56: 1532-1536
33. Chang CT, Wu CSC, Yang JT. Circular dichroic analysis of protein conformation: inclusion of the β-turns. Anal Biochem,1978,91:13-31
34. Chen JS, Sappington TW, Raikhel AS. Extensive sequence conservation among insect, nematode, and vertebrate vitellogenins reveals ancient common ancestry. J Mol Evol, 1997, 44: 440-451
35. Chen PS, Toribara TY, Warner H. Microdetermination of phosphorus. Anal Chem, 1956,28:1756-1758
36. Chen Y, Bal B, Gorski J. Calcium and collagen binding properties of osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 from bone. JBiol Chem, 1992, 267:24871-24878
37. Choi I, Jung C, Choi H, Kim C, Ha H. Effectiveness of phosvitin peptides on enhancing bioavailability of calcium and its accumulation in bones. Food Chem, 2005,93:577-583
38. Choi I, Jung C, Seog H, Choi H. Purification of phosvitin from egg yolk and determination of its physicochemical properties. Food Sci Biotechnol, 2004, 13: 434-437
39. Christensen B, Nielsen Mxas, Haselmann Kxaf, Petersen Txae, Sorensen E. Post-translationally modified residues of native human osteopontin are located in clusters: identification of 36 phosphorylation and five O-glycosylation sites and their biological implications. Biochem J, 2005,390: 285-292
40. Chu X, Jiang W, Zhang Z, Yan Y, Pan H, Xu X, Tang R. Unique roles of acidic amino acids in phase transformation of calcium phosphates. JPhys Chem B, 2010, 115:1151-1157
41. Chung SL, Ferrier LK. pH and sodium chloride effects on emulsifying properties of egg yolk phosvitin. J Food Sci, 1992, 57:40-42
42. Chung SL, Ferrier LK. Heat denaturation and emulsifying properties of egg yolk phosvitin. J Food Sci, 1995, 60:906-908
43. Clark R. The isolation and composition of two phosphoproteins from hen's egg. Biochem J, 1970, 118:537-542
44. Clark RC. Amino acid sequence of a cyanogen bromide cleavage peptide from hens egg phosvitin. Biochim Biophys Acta, 1973,310:174-187
45. Clark RC. The primary structure of avian phosvitins:Contributions through the edman degradation of methylmercaptovitins prepared from the constituent phosphoproteins. Int J Biochem, 1985,17:983-988
46. Conner SD, Schmid SL. Regulated portals of entry into the cell. Nature, 2003,422: 37-44
47. Cookson DJ, Levine BA, Williams RJP, Jontell M, Linde A, Bernard B. Cation binding by the Rat-incisor-dentine phosphoprotein. Eur J Biochem, 1980, 110: 273-278
48. Curtin P, McHugh KP, Zhou H-Y, Fluckiger R, Goldhaber P, Oppenheim FG, Salih E. Modulation of bone resorption by phosphorylation state of bone sialoprotein. Biochemistry, 2009,48:6876-6886
49. Cutting JA, Roth TF. Staining of phospho-proteins on acrylamide gel electropherograms. Anal Biochem, 1973,54:386-394
50. Czernick D, Liu J, Serge D, Salih E. Topographical distribution of phosphorylation sites of phosvitins by mass spectrometry. J Proteomics, 2013,83: 76-98
51. Dahlin S, Angstrom J, Linde A. Dentin phosphoprotein sequence motifs and molecular modeling: conformational adaptations to mineral crystals. Eur J Oral Sci,1998,106:239-248
52. Damodaran S, Xu S. The role of electrostatic forces in anomalous adsorption behavior of phosvitin at the air/water interface. J Colloid Interface Sci, 1996, 178: 426-435
53. Dorozhkin SV. Nanodimensional and nanocrystalline calcium orthophosphates. Biomed. Mater. Diagn. Devices, 2012:19
54. Doucet D, Otter DE, Gauthier SF, Foegeding EA. Enzyme-induced gelation of extensively hydrolyzed whey proteins by Alcalase: peptide identification and determination of enzyme specificity. JAgric Food Chem, 2003,51:6300-6308
55. Dunker AK, Brown CJ, Lawson JD, Iakoucheva LM, Obradovic Z. Intrinsic disorder and protein function. Biochemistry, 2002, 41:6573-6582
56. Feng F, Mine Y. Phosvitin phosphopeptides increase iron uptake in a Caco-2 cell monolayer model. Int J Food Sci Technol, 2006, 41: 455-458
57. Ferraretto A, Gravaghi C, Fiorilli A, Tettamanti G. Casein-derived bioactive phosphopeptides:role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells. FEBS Lett, 2003,551:92-98
58. Finn RN. Vertebrate yolk complexes and the functional implications of phosvitins and other subdomains in vitellogenins. Biol. Reprod,2007,76:926-935
59. Finn RN, Kristoffersen BA. Vertebrate vitellogenin gene duplication in relation to the "3R hypothesis": correlation to the pelagic egg and the oceanic radiation of teleosts. PLoS One, 2007,2:e169
60. Finn RN, Ostby GC, Norberg B, Fyhn HJ. In vivo oocyte hydration in Atlantic halibut (Hippoglossus hippoglossus); proteolytic liberation of free amino acids, and ion transport, are driving forces for osmotic water influx. J Exp Biol, 2002, 205:211-224
61. Fologea D, Ledden B, McNabb DS, Li J. Electrical characterization of protein molecules by a solid-state nanopore. Appl Phys Lett, 2007, 91:053901-053903
62. Francis G, Fisher D, Delgado C, Malik F, Gardiner A, Neale D. PEGylation of cytokines and other therapeutic proteins and peptides: the importance of biological optimisation of coupling techniques. Int J Hematol, 1998, 68:1-18
63. Geladopoulos TP, Sotiroudis TG. Interaction of aluminum ions with phosvitin. J Inorg Biochem,1994,54: 247-256
64. George A, Veis A. Phosphorylated proteins and control over apatite nucleation, crystal growth, and inhibition. Chem Rev, 2008, 108:4670-4693
65. Gericke A, Qin C, Spevak L, Fujimoto Y, Butler W, Sorensen ES, Boskey A. Importance of phosphorylation for osteopontin regulation of biomineralization. Calcif Tissue Int, 2005, 77:45-54
66. Glimcher MJ. Bone:Nature of the calcium phosphate crystals and cellular, structural, and physical chemical mechanisms in their formation. Rev Mineral Geochem,2006,64:223-282
67. Goulas A, Triplett EL, Taborsky G. Oligophosphopeptides of varied structural complexity derived from the egg phosphoprotein, phosvitin. Protein J, 1996, 15: 1-9
68. Greengard O, Sentenac A, Mendelsohn N. Phosvitin, the iron carrier of egg yolk. Biochim Biophys Acta, 1964, 90: 406-407
69. Grizzuti K, Perlmann GE. Conformation of the phosphoprotein, phosvitin. J Biol Chem,1970,245:2573-2578
70. Grizzuti K, Perlmann GE. Binding of magnesium and calcium ions to the phosphoglycoprotein phosvitin. Biochemistry, 1973,12:4399-4403
71. Grizzuti K, Perlmann GE. Further studies on the binding of divalent cations to the phosphoglycoprotein phosvitin. Biochemistry, 1975,14:2171-2175
72. Grogan J, Taborsky G. Iron binding by phosvitin: variation of rate of iron release as a function of the degree of saturation of iron binding sites. J Inorg Biochem, 1986,26:229-236
73. Grohe B, O'Young J, Ionescu DA, Lajoie G, Rogers KA, Karttunen M, Goldberg HA, Hunter GK. Control of calcium oxalate crystal growth by face-specific adsorption of an osteopontin phosphopeptide. J Am Chem Soc, 2007, 129: 14946-14951
74. Guerin-Dubiard C, Anton M, Dhene-Garcia A, Martinet V, Brule G. Hen egg and fish egg phosvitins: composition and iron binding properties. Eur Food Res Technol, 2002, 214:460-464
75. Honig W, Kula M-R. Selectivity of protein precipitation with polyethylene glycol fractions of various molecular weights. Anal Biochem, 1976, 72:502-512
76. He G, Dahl T, Veis A, George A. Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1. Nat Mater, 2003,2:552-558
77. He G, Ramachandran A, Dahl T, George S, Schultz D, Cookson D, Veis A, George A. Phosphorylation of phosphophoryn is crucial for its function as a mediator of biomineralization. JBiol Chem,2005,280:33109-33114
78. Hegenauer J, Saltman P, Nace G. Iron(III)-phosphoprotein chelates: Stoichiometric equilibrium constant for interaction of iron(iii) and phosphorylserine residues of phosvitin and casein. Biochemistry, 1979, 18: 3865-3879
79. Hiramatsu N, Cheek AO, Sullivan CV, Matsubara T, Hara A. Vitellogenesis and endocrine disruption. Biochem Mol Biol Fishes, 2005,6:431-471
80. Hiramatsu N, Ichikawa N, Fukada H, Fujita T, Sullivan CV, Hara A. Identification and characterization of proteases involved in specific proteolysis of vitellogenin and yolk proteins in salmonids. JExp Zool, 2002, 292:11-25
81. Hiramatsu N, Matsubara T, Fujita T, Sullivan CV, Hara A. Multiple piscine vitellogenins:biomarkers of fish exposure to estrogenic endocrine disruptors in aquatic environments. Mar Biol, 2006, 149: 35-47
82. Huang H-D, Lee T-Y, Tzeng S-W, Horng J-T. KinasePhos:a web tool for identifying protein kinase-specific phosphorylation sites. Nucleic Acids Res, 2005, 33:W226-W229
83. Hunter G, Hauschka P, Poole A, Rosenberg L, Goldberg H. Nucleation and inhibition of hydroxyapatite formation by mineralized tissue proteins. Biochem J, 1996,317:59-64
84. Hunter GK, Goldberg HA. Modulation of crystal formation by bone phosphoproteins:role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein. Biochem J, 1994, 302:175-179
85. Huq NL, Cross KJ, Ung M, Reynolds EC. A review of protein structure and gene organisation for proteins associated with mineralised tissue and calcium phosphate stabilisation encoded on human chromosome 4. Arch Oral Biol, 2005, 50: 599-609
86. Ibanez A, Peinado-Onsurbe J, Sanchez E, Prat F. The role of lipoprotein lipase (LPL) in the incorporation of neutral lipids into the oocytes of the European sea bass (Dicentrarchus labrax L.) during gonadal development. Fish Physiol Biochem, 2003,28:291-293
87. Ingham KC. Precipitation of proteins with polyethylene glycol. Method Enzymol, 1989,182:301-306
88. Ishikawa SI, Yano Y, Arihara K, Itoh M. Egg yolk phosvitin inhibits hydroxyl radical formation from the fenton reaction. Biosci, Biotechnol, Biochem, 2004, 68: 1324-1331
89. Ito S, Iijima M, Motai F, Mizoguchi I, Saito T. Effects of calcium salts of acidic monomers on mineral induction of phosphoprotein immobilized to agarose beads. J Biomed Mater Res Part A,2012,100:2760-2765
90. Itoh T, Abe Y, Adachi S. Comparative studies on the alpha and beta-phosvitin from hen's egg yolk. J Food Sci, 1983,48:1755-1757
91. Jiang B, Mine Y. Preparation of novel functional oligophosphopeptides from hen egg yolk phosvitin. JAgric Food Chem, 2000,48:990-994
92. Jiang B, Mine Y. Phosphopeptides derived from hen egg yolk phosvitin: effect of molecular size on the calcium-binding properties. Biosci, Biotechnol, Biochem, 2001,65:1187-1190
93. Joubert FJ, Cook WH. Preparation and characterization of phosvitin from hen egg yolk. Can JBiochem Physiol, 1958, 36:399-408
94. Jung S, Kim DH, Son JH, Nam K, Ahn DU, Jo C. The functional property of egg yolk phosvitin as a melanogenesis inhibitor. Food Chem, 2012, 135:993-998
95. Karampas IA, Kontoyannis CG. Characterization of calcium phosphates mixtures. Vib Spectrosc,2013,64:126-133
96. Kato A, Miyazaki S, Kawamoto A, KOBAYASHI K. Effects of phosphate residues on the excellent emulsifying properties of phosphoglycoprotein phosvitin (food & nutrition). Agric Biol Chem,1987,51:2989-2994
97. Keykhosravani M, Doherty-Kirby A, Zhang C, Brewer D, Goldberg HA, Hunter GK, Lajoie G. Comprehensive identification of post-translational modifications of rat bone osteopontin by mass spectrometry. Biochemistry, 2005,44: 6990-7003
98. Khan MAS, Nakamura S, Ogawa M, Akita E, Azakami H, Kato A. Bactericidal action of egg yolk phosvitin against Escherichia coli under thermal stress. J Agric Food Chem,2000,48:1503-1506
99. Ko KY, Nam KC, Jo C, Lee EJ, Ahn DU. A simple and efficient method for preparing partially purified phosvitin from egg yolk using ethanol and salts. Poultry Sci, 2011,90:1096-1104
100.Kobayashi N, Onuma K, Oyane A, Yamazaki A. The role of phosvitin for nucleation of calcium phosphates on collagen. Key Eng Mater, 2004, 254-256: 537-540
101. Kozlowski H, Mangani S, Messori L, Orioli PL, Scozzafava A. CD and EXAFS study of the interaction between phosvitin and copper(II) ions. J Inorg Biochem, 1988,34: 221-239
102. Kumar V, Sharma VK, Kalonia DS. Effect of polyols on polyethylene glycol (PEG)-induced precipitation of proteins:Impact on solubility, stability and conformation. Int J Pharm,2009,366:38-43
103. Kwak S-Y, Wiedemann-Bidlack FB, Beniash E, Yamakoshi Y, Simmer JP, Litman A, Margolis HC. Role of 20-kDa amelogenin (P148) phosphorylation in calcium phosphate formation in vitro. JBiol Chem, 2009, 284: 18972-18979
104. Lea C, Hawke J. Lipovitellin. 2. The influence of water on the stability of lipovitellin and the effects of freezing and of drying. Biochem J, 1952, 52: 105-114
105. Lee SK, Han JH, Decker EA. Antioxidant activity of phosvitin in phosphatidylcholine liposomes and meat model systems. J Food Sci, 2002, 67: 37-41
106. Lee SL, Veis A, Glonek T. Dentin phosphoprotein: an extracellular calcium-binding protein. Biochemistry, 1977, 16:2971-2979
107. Lei B, Wu J. Purification of egg yolk phosvitin by anion exchange chromatography. JChromatogr A,2012, 1223:41-46
108. Levene P, Alsberg C. Zur Chemie der Paranucleinsaure. Hoppe-Seyler's Zeitschrift fur physiologische Chemie, 1901,31:543-555
109. Li B, Raff J, Barkleit A, Bernhard G, Foerstendorf H. Complexation of U(VI) with highly phosphorylated protein, phosvitin. A vibrational spectroscopic approach. J Inorg Biochem,2010,104: 718-725
110. Lim EH, Teo BY, Lam TJ, Ding JL. Sequence analysis of a fish vitellogenin cDNA with a large phosvitin domain. Gene, 2001,277:175-186
111. Linde A, Lussi A, Crenshaw M. Mineral induction by immobilized polyanionic proteins. Calcif Tissue Int,1989, 44: 286-295
112. Lipmann FA, Levene P. Serinephosphoric acid obtained on hydrolysis of vitellinic acid. JBiol Chem,1932,98:109-114
113. Llena C, Forner L, Baca P. Anticariogenicity of casein phosphopeptide-amorphous calcium phosphate: a review of the literature. J Contemp Dent Pract, 2009, 10:1-9
114.Losso J, Nakai S. A simple procedure for the isolation of phosvitin from chicken egg yolk. In: ed. Egg Uses and Processing Technologies:New Developments. Oxon, UK: Cab International,1994:150-157
115. Lu C-L, Baker RC. Effect of pH and Food Ingredients on the Stability of Egg Yolk Phospholipids and the Metal-Chelator Antioxidant Activity of Phosvitin. J Food Sci,1987,52:613-616
116. Ma J, Wang H, Wang Y, Zhang S. Endotoxin-neutralizing activity of hen egg phosvitin. Mol Immunol,2013,53:355-362
117. Mann S. Mineralization in biological systems. In: ed. Inorganic Elements in Biochemistry: Springer Berlin Heidelberg, 1983:125-174
118. Mann S. Molecular tectonics in biomineralization and biomimetic materials chemistry. Nature, 1993,365:499-505
119. Matsudomi N, Ito K, Yoshika Y. Preventive effect of egg yolk phosvitin on heat-insolubilization of egg white protein and its application to heat-induced egg white gel. Biosci, Biotechnol, Biochem, 2006, 70:836-842
12O. Mecham DK, Olcott HS. Phosvitin, the principal phosphoprotein of egg yolk. J Am Chem Soc,1949,71:3670-3679
121. Meisel H, Meisel H, Fairweather-Tait S, FitzGerald RJ, Hartmann R, Lane C, McDonagh D, Teucher B, Wal J. Detection of caseinophosphopeptides in the distal ileostomy fluid of human subjects. Brit J Nutr, 2003,89:351-358
122. Mikroulis D, Mavrilas D, Kapolos J, Koutsoukos P, Lolas C. Physicochemical and microscopical study of calcific deposits from natural and bioprosthetic heart valves. Comparison and implications for mineralization mechanism. J Mater Sci: Mater Med, 2002, 13:885-889
123. Mine Y. Egg bioscience and biotechnology: John Wiley & Sons, 2008:42-47
124. Moran E. Nutrition of the developing embryo and hatchling. Poultry Sci, 2007, 86: 1043-1049
125. Mosconi G, Carnevali O, Habibi H, Sanyal R, Polzonetti-Magni A. Hormonal mechanisms regulating hepatic vitellogenin synthesis in the gilthead sea bream, Sparus aurata. Am JPhysiol-cell Ph, 2002, 283:C673-C678
126. Nakamura S, Ogawa M, Nakai S, Kato A, Kitts DD. Antioxidant activity of a Maillard-type phosvitin-galactomannan conjugate with emulsifying properties and heat stability. J Agric Food Chem,1998, 46: 3958-3963
127. Obenauer JC, Cantley LC, Yaffe MB. Scansite 2.0: Proteome-wide prediction of cell signaling interactions using short sequence motifs. Nucleic Acids Res, 2003, 31:3635-3641
128. Onuma K. Effect of phosvitin on the nucleation and growth of calcium phosphates in physiological solutions. JPhys Chem B, 2005, 109:8257-8262
129. Osaki S, Sexton RC, Pascual E, Frieden E. Iron oxidation and transferrin formation by phosvitin. Biochem J, 1975, 151:519-525
130. Oshiro M, Yamaguchi K, Takamizawa T, Inage H, Watanabe T, Irokawa A, Ando S, Miyazaki M. Effect of CPP-ACP paste on tooth mineralization: an FE-SEM study. Journal of Oral Science, 2007, 49: 115-120
131. Palmer LC, Newcomb CJ, Kaltz SR, Spoerke ED, Stupp SI. Biomimetic systems for hydroxyapatite mineralization inspired by bone and enamel. Chem Rev, 2008, 108:4754-4783
132. Pan M, Bell WJ, Telfer WH. Vitellogenic blood protein synthesis by insect fat body. Science, 1969, 165:393-394
133. Pasupuleti VK, Braun S. State of the art manufacturing of protein hydrolysates. In: ed. Protein Hydrolysates in Biotechnology: Springer Netherlands, 2010:11-32
134. Perlmann GE, Grizzuti K. Conformational transition of the phosphoprotein phosvitin. Random conformation-β structure. Biochemistry, 1971,10:258-264
135. Prescott B, Renugopalakrishnan V, Glimcher MJ, Bhushan A, Thomas Jr GJ. A Raman spectroscopic study of hen egg yolk phosvitin: structures in solution and in the solid state. Biochemistry,1986,25:2792-2798
136. Raldua D, Fabra M, Bozzo MG, Weber E, Cerda J. Cathepsin B-mediated yolk protein degradation during killifish oocyte maturation is blocked by an H+-ATPase inhibitor: effects on the hydration mechanism. Am J Physiol: Regul, Integr Comp Physiol, 2006, 290: R456-R466
137. Reese C, Heise F, Mayer A. Trans-SNARE pairing can precede a hemifusion intermediate in intracellular membrane fusion. Nature,2005,436:410-414
138. Ren J. Phosvitin extraction and phosphopeptides characterization from chicken egg yolk [Master's thesis]. Edmonton: University of Alberta Libraries, 2012.
139. Renugopalakrishnan V, Horowitz PM, Glimcher MJ. Structural studies of phosvitin in solution and in the solid state. JBiol Chem 1985, 260: 11406-11413
140.Reynolds E. Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions. J Dent Res, 1997, 76: 1587-1595
141. Reynolds EC. Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: a review. Spec Care Dentist, 1998, 18:8-16
142. Romano M, Rosanova P, Anteo C, Limatola E. Vertebrate yolk proteins: A review. Mol Reprod Dev, 2004, 69:109-116
143. Saad FA, Salih E, Glimcher MJ. Identification of osteopontin phosphorylation sites involved in bone remodeling and inhibition of pathological calcification. J Cell Biochem,2008,103:852-856
144. Saito T, Arsenault AL, Yamauchi M, Kuboki Y, Crenshaw MA. Mineral induction by immobilized phosphoproteins. Bone, 1997, 21:305-311
145. Salih E, Wang J, Mah J, Fluckiger R. Natural variation in the extent of phosphorylation of bone phosphoproteins as a function of in vivo new bone formation induced by demineralized bone matrix in soft tissue and bony environments. Biochem J, 2002, 364:465-474
146. Salih E, Zhou H-Y, Glimcher MJ. Phosphorylation of purified bovine bone sialoprotein and osteopontin by protein kinases. J Biol Chem, 1996, 271: 16897-16905
147. Samaraweera H, Zhang WG, Lee EJ, Ahn DU. Egg yolk phosvitin and functional phosphopeptides-Review. J Food Sci, 2011,76: R143-R150
148. Samaraweera Mudiyanselage CH. Production and characterization of phosphopeptides from egg yolk phosvitin [Ph D dissertation]. Ames:Iowa State University Libraries, 2012.
149. Sappington TW, Raikhel AS. Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem Mol Biol, 1998, 28:277-300
150. Selman K, Wallace RA, Cerda J. Bafilomycin A1 inhibits proteolytic cleavage and hydration but not yolk crystal disassembly or meiosis during maturation of sea bass oocytes. JExp Zool, 2001, 290:265-278
151. Sen LC, Gonzalez-Flores E, Feeney RE, Whitaker JR. Reactions of phosphoproteins in alkaline solutions. JAgric Food Chem, 1977, 25:632-638
152. Shainkin R, Perlmann GE. Phosvitin, a phosphoglycoprotein. I. Isolation and characterization of a glycopeptide from phosvitin. J Biol Chem, 1971a, 246: 2278-2284
153. Shainkin R, Perlmann GE. Phosvitin, a phosphoglycoprotein: Composition and partial structure of carbohydrate moiety. Arch Biochem Biophys, 1971b, 145: 693-700
154. Sharrock WJ, Rosenwasser TA, Gould J, Knott J, Hussey D, Gordon JI, Banaszak L. Sequence of lamprey vitellogenin. Implications for the lipovitellin crystal structure. Journal of Molecular Biology, 1992, 226: 903-907
155. Shkilnyy A, Friedrich A, Tiersch B, Schone S, Fechner M, Koetz J, Schlapfer C-W, Taubert A. Poly (ethylene imine)-controlled calcium phosphate mineralization. Langmuir, 2008,24:2102-2109
156. Shkilnyy A, Schone S, Rumplasch C, Uhlmann A, Hedderich A, Giinter C, Taubert A. Calcium phosphate mineralization with linear poly (ethylene imine): a time-resolved study. Colloid Polym Sci, 2011,289:881-888
157. Silva R, Fischer A, Burch J. The major and minor chicken vitellogenin genes are each adjacent to partially deleted pseudogene copies of the other Mol Cell Biol, 1989,9:3557-3562
158. Silversand C, Haux C. Fatty acid composition of vitellogenin from four teleost species. Journal of Comparative Physiology B, 1995, 164:593-599
159. Sobota M, Salejda A, Trziszka T. Effect of the phosvitin isolation metod on yield of the process. In: 3rd Baltic Conference on Food Science and Technology FOODBALT-2008. Jelgava; 2008:148-151.
160. Spackman DH, Stein WH, Moore S. Automatic recording apparatus for use in chromatography of amino acids. Anal Chem, 1958, 30:1190-1206
161. Sreenath T, Thyagarajan T, Hall B, Longenecker G, D'Souza R, Hong S, Wright JT, MacDougall M, Sauk J, Kulkarni AB. Dentin sialophosphoprotein knockout mouse teeth display widened predentin zone and develop defective dentin mineralization similar to human dentinogenesis imperfecta type III. J Biol Chem, 2003,278:24874-24880
162. Stetler-Stevenson W, Veis A. Bovine dentin phosphophoryn: calcium ion binding properties of a high molecular weight preparation. Calcif Tissue Int, 1987, 40: 97-102
163. Sun C, Hu L, Liu S, Hu G, Zhang S. Antiviral activity of phosvitin from zebrafish Danio rerio. Dev Comp Immunol, 2013, 40:28-34
164. Sundararajan TA, Sampath Kumar KSV, Sarma PS. A simplified procedure for the preparation of phosvitin and vitellin. Biochim Biophys Acta, 1960, 38:360-362
165. Sutton RB, Fasshauer D, Jahn R, Brunger AT. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature, 1998, 395: 347-354
166. Taborsky G. Interaction between phosvitin and iron and its effect on a rearrangement of phosvitin structure. Biochemistry, 1963, 2:266-271
167. Taborsky G. Optical rotatory dispersion and circular dichroism of phosvitin at low pH. Reversible transition between unordered conformation and beta structure. J Biol Chem,1968,243:6014-6020
168. Taborsky G. Effect of freezing and thawing on the conformation of phosvitin. J Biol Chem,1970,245:1054-1062
169. Taborsky G. Iron binding by phosvitin and its conformational consequences. J Biol Chem,1980,255:2976-2984
170. Taborsky G. Phosvitin. Advances in inorganic biochemistry, 1983,5:235-279
171. Taborsky G. On the interaction of phosvitins with ferric ion: Solubility of the Fe(III)-phosphoprotein complex under acidic conditions is a function of the iron/phosphate ratio and the degree of phosvitin phosphorylation. J Inorg Biochem, 1991,44:65-77
172. Taborsky G, Allende CC. A rearrangement in the structure of phosvitin. Biochemistry, 1962,1:406-411
173. Taborsky G, Mok CC. Phosvitin. Homogeneity and molecular weight. J Biol Chem,1967,242:1495-1501
174. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 2011,28:2731-2739
175. Tay F, Pashley D. Biomimetic remineralization of resin-bonded acid-etched dentin. J Dent Res,2009,88:719-724
176. Ternes W. Characterization of water soluble egg yolk proteins with isoelectric focusing. J Food Sci, 1989, 54: 764-765
177. Thompson JR, Banaszak LJ. Lipid-protein interactions in lipo vitellin. Biochemistry,2002,41:9398-9409
178. Veis A. Mineralization in organic matrix frameworks. Reviews in Mineralogy and Geochemistry, 2003, 54: 249-289
179. Vogel HJ. Structure of hen phosvitin: A 31P NMR, 1H NMR, and laser photochemically induced dynamic nuclear polarization 1H NMR study. Biochemistry, 1983,22:668-674
18O. Wallace RA, Morgan JP. Chromatographic resolution of chicken phosvitin. Multiple macromolecular species in a classic vitellogenin-derived phosphoprotein. Biochem J, 1986a, 240:871-878
181. Wallace RA, Morgan JP. Isolation of phosvitin: Retention of small molecular weight species and staining characteristics on electrophoretic gels. Anal Biochem, 1986b,157: 256-261
182. Wallace RA, Selman K. Ultrastructural aspects of oogenesis and oocyte growth in fish and amphibians. J Electron Microsc Tech, 1990, 16:175-201
183. Wallwork M, Kirkham J, Chen H, Chang S, Robinson C, Smith D, Clarkson B. Binding of dentin noncollagenous matrix proteins to biological mineral crystals: an atomic force microscopy study. Calcif Tissue Int, 2002, 71:249-256
184. Wang S-Y, Williams D. Biosynthesis of the vitellogenins. Identification and characterization of nonphosphorylated precursors to avian vitellogenin I and vitellogenin Ⅱ. JBiol Chem, 1982, 257:3837-3846
185. Wang S, Smith DE, Williams DL. Purification of avian vitellogenin III: comparison with vitellogenins I and II. Biochemistry, 1983,22: 6206-6212
186. Wang S, Wang Y, Ma J, Ding Y, Zhang S. Phosvitin plays a critical role in the immunity of zebrafish embryos via acting as a pattern recognition receptor and an antimicrobial effector. JBiol Chem,2011,286:22653-22664
187. Wang SY, Williams DL. Identification, purification, and characterization of two distinct avian vitellogenins. Biochemistry, 1980, 19:1557-1563
188. Wiedemann-Bidlack FB, Kwak S-Y, Beniash E, Yamakoshi Y, Simmer JP, Margolis HC. Effects of phosphorylation on the self-assembly of native full-length porcine amelogenin and its regulation of calcium phosphate formation in vitro. J Struct Biol, 2011,173:250-260
189. Xiao S, Yu C, Chou X, Yuan W, Wang Y, Bu L, Fu G, Qian M, Yang J, Shi Y. Dentinogenesis imperfecta 1 with or without progressive hearing loss is associated with distinct mutations in DSPP. Nat Genet, 2001, 27:201-204
19O. Yamamura JI, Adachi T, Aoki N, Nakajima H, Nakamura R, Matsuda T. Precursor-product relationship between chicken vitellogenin and the yolk proteins: The 40 kDa yolk plasma glycoprotein is derived from the C-terminal cysteine-rich domain of vitellogenin II. Biochim Biophys Acta, 1995, 1244: 384-394
191. Yasui SC, Pancoska P, Dukor RK, Keiderling TA, Renugopalakrishnan V, Glimcher MJ, Clark RC. Conformational transitions in phosvitin with pH variation. Vibrational circular dichroism study. JBiol Chem, 1990, 265:3780-3788
192. Yoshizaki N, Yonezawa S. Cathepsin D activity in the vitellogenesis of Xenopus laevis. Dev, Growth Differ, 1994,36:299-306
193. Young D, Nau F, Pasco M, Mine Y. Identification of hen egg yolk-derived phosvitin phosphopeptides and their effects on gene expression profiling against oxidative stress-induced caco-2 cells. JAgric Food Chem, 2011,59:9207-9218
194. Zachariou M, Traverso I, Spiccia L, Hearn MT. Potentiometric investigations into the acid-base and metal ion binding properties of immobilized metal ion affinity chromatographic (IMAC) adsorbents. JPhys Chem, 1996, 100:12680-12690
195. Zhang X, Qiu N, Geng F, Ma M. Simply and effectively preparing high-purity phosvitin using polyethylene glycol and anion-exchange chromatography. J Sep Sci,2011,34:3295-3301
196. Znidarsic WJ, Chen IW, Shastri VP. Influence of surface charge and protein intermediary layer on the formation of biomimetic calcium phosphate on silica nanoparticles. J Mater Chem, 2012, 22:19562-19569
2.倪莉,王璋,许时婴.鸡蛋中卵黄高磷蛋白的提取.无锡轻工大学学报，1999：57-61
3.徐彩娜.卵黄高磷蛋白磷酸肽的抗炎机理研究[博士学位论文].长春：吉林大学,2012
4. Abdelkebir K, Morin-Grognet S, Gaudiere F, Coquerel G, Labat B, Atmani H, Ladam G. Biomimetic layer-by-layer templates for calcium phosphate biomineralization. Acta Biomater, 2012,8:3419-3428
5. Abe Y, Itoh T, Adachi S. Fractionation and characterization of hen's egg yolk phosvitin. J Food Sci, 1982, 47: 1903-1907
6. Albright K, Gordon D, Cotterill O. Release of iron from phosvitin by heat and food additives. J Food Sci,1984,49:78-81
7. Allerton SE, Perlmann GE. Chemical characterization of the phosphoprotein phosvitin. JBiol Chem, 1965, 240: 3892-3898
8. Anton M. Composition and structure of hen egg yolk. In: ed. Bioactive egg compounds:Springer Berlin Heidelberg, 2007:1-6
9. Anton M, Castellani O, Guerin-Dubiard C. Phosvitin. In: ed. Bioactive Egg Compounds:Springer Berlin Heidelberg, 2007:17-24
10. Arifuzzaman S, Rohani S. Experimental study of brushite precipitation. J Cryst Growth,2004,267:624-634
11. Atha DH, Ingham KC. Mechanism of precipitation of proteins by polyethylene glycols. Analysis in terms of excluded volume. JBiol Chem, 1981,256:12108
12. Baht G, O'Young J, Borovina A, Chen H, Tye C, Karttunen M, Lajoie G, Hunter G, Goldberg H. Phosphorylation of Ser136 is critical for potent bone sialoprotein-mediated nucleation of hydroxyapatite crystals. Biochem J, 2010, 428: 385-395
13. Bassani G, Farruggia B, Nerli B, Romanini D, Pico G. Porcine pancreatic lipase partition in potassium phosphate-polyethylene glycol aqueous two-phase systems. J Chromatogr B, 2007, 859:222-228
14. Bee L, Cotterill O. Ion-exchange chromatography and electrophoresis of egg yolk proteins. J Food Sci, 1979,44:656-667
15. Belhomme C, David-Briand E, Guerin-Dubiard C, Vie V, Anton M. Phosvitin-calcium aggregation and organization at the air-water interface. Colloids Surf, B, 2008,63:12-20
16. Belhomme C, David-Briand E, Ropers M-H, Guerin-Dubiard C, Anton M. Interfacial characteristics of spread films of hen egg yolk phosvitin at the air-water interface: Interrelation with its charge and aggregation state. Food Hydrocolloids,2007,21:896-905
17. Bos MA, Vliet Tv. Interfacial rheological properties of adsorbed protein layers and surfactants: a review. Adv Colloid Interface Sci, 2001,91:437-471
18. Boskey AL. Mineral-matrix interactions in bone and cartilage. Clin Orthop Relat R,1992,281:244-274
19. Boskey AL. Matrix proteins and mineralization: an overview. Connect Tissue Res, 1996,35:357-363
20. Brockbank RL, Vogel HJ. Structure of the oligosaccharide of hen phosvitin as determined by two-dimensional 1H NMR of the intact glycoprotein. Biochemistry, 1990,29:5574-5583
21. Burke E, Guo Y, Colon L, Rahima M, Veis A, Nancollas G. Influence of polyaspartic acid and phosphophoryn on octacalcium phosphate growth kinetics. Colloids Surf, B, 2000, 17:49-57
22. Burley RW, Cook WH. Isolation and composition of avian egg yolk granules and their constituent α-and β-lipovitellins. Can J Biochem Physiol, 1961,39: 1295-1307
23. Byrne BM, Gruber M, Ab G. The evolution of egg yolk proteins. Prog Biophys Mol Bio,1989,53:33-69
24. Byrne BM, Van het Schip AD, Van de Klundert JA, Arnberg AC, Gruber M, Ab G. Amino acid sequence of phosvitin derived from the nucleotide sequence of part of the chicken vitellogenin gene. Biochemistry, 1984, 23:4275-4279
25. Carbonaro M, Nucara A. Secondary structure of food proteins by Fourier transform spectroscopy in the mid-infrared region. Amino Acids, 2010, 38: 679-690
26. Carnevali O, Cionna C, Tosti L, Lubzens E, Maradonna F. Role of cathepsins in ovarian follicle growth and maturation. Gen Comp Endocr, 2006, 146:195-203
27. Castellani O, Belhomme C, David-Briand E, Guerin-Dubiard C, Anton M. Oil-in-water emulsion properties and interfacial characteristics of hen egg yolk phosvitin. Food Hydrocolloids, 2006, 20:35-43
28. Castellani O, Belhomme C, David-Briand E, Guerin-Dubiard C, Anton M. The role of metal ions in emulsion characteristics and flocculation behaviour of phosvitin-stabilised emulsions. Food Hydrocolloids, 2008, 22:1243-1253
29. Castellani O, David-Briand E, Guerin-Dubiard C, Anton M. Effect of aggregation and sodium salt on emulsifying properties of egg yolk phosvitin. Food Hydrocolloids,2005,19:769-776
30. Castellani O, Guerin-Dubiard C, David-Briand E, Anton M. Influence of physicochemical conditions and technological treatments on the iron binding capacity of egg yolk phosvitin. Food Chem, 2004,85:569-577
31. Castellani O, Martinet V, David-Briand E, Guerin-Dubiard C, Anton M. Egg yolk phosvitin: Preparation of metal-free purified protein by fast protein liquid chromatography using aqueous solvents. J Chromatogr B, 2003,791:273-284
32. Causeret D, Matringe E, Lorient D. Ionic strength and pH effects on composition and microstructure of yolk granules. JFood Sci, 1991,56: 1532-1536
33. Chang CT, Wu CSC, Yang JT. Circular dichroic analysis of protein conformation: inclusion of the β-turns. Anal Biochem,1978,91:13-31
34. Chen JS, Sappington TW, Raikhel AS. Extensive sequence conservation among insect, nematode, and vertebrate vitellogenins reveals ancient common ancestry. J Mol Evol, 1997, 44: 440-451
35. Chen PS, Toribara TY, Warner H. Microdetermination of phosphorus. Anal Chem, 1956,28:1756-1758
36. Chen Y, Bal B, Gorski J. Calcium and collagen binding properties of osteopontin, bone sialoprotein, and bone acidic glycoprotein-75 from bone. JBiol Chem, 1992, 267:24871-24878
37. Choi I, Jung C, Choi H, Kim C, Ha H. Effectiveness of phosvitin peptides on enhancing bioavailability of calcium and its accumulation in bones. Food Chem, 2005,93:577-583
38. Choi I, Jung C, Seog H, Choi H. Purification of phosvitin from egg yolk and determination of its physicochemical properties. Food Sci Biotechnol, 2004, 13: 434-437
39. Christensen B, Nielsen Mxas, Haselmann Kxaf, Petersen Txae, Sorensen E. Post-translationally modified residues of native human osteopontin are located in clusters: identification of 36 phosphorylation and five O-glycosylation sites and their biological implications. Biochem J, 2005,390: 285-292
40. Chu X, Jiang W, Zhang Z, Yan Y, Pan H, Xu X, Tang R. Unique roles of acidic amino acids in phase transformation of calcium phosphates. JPhys Chem B, 2010, 115:1151-1157
41. Chung SL, Ferrier LK. pH and sodium chloride effects on emulsifying properties of egg yolk phosvitin. J Food Sci, 1992, 57:40-42
42. Chung SL, Ferrier LK. Heat denaturation and emulsifying properties of egg yolk phosvitin. J Food Sci, 1995, 60:906-908
43. Clark R. The isolation and composition of two phosphoproteins from hen's egg. Biochem J, 1970, 118:537-542
44. Clark RC. Amino acid sequence of a cyanogen bromide cleavage peptide from hens egg phosvitin. Biochim Biophys Acta, 1973,310:174-187
45. Clark RC. The primary structure of avian phosvitins:Contributions through the edman degradation of methylmercaptovitins prepared from the constituent phosphoproteins. Int J Biochem, 1985,17:983-988
46. Conner SD, Schmid SL. Regulated portals of entry into the cell. Nature, 2003,422: 37-44
47. Cookson DJ, Levine BA, Williams RJP, Jontell M, Linde A, Bernard B. Cation binding by the Rat-incisor-dentine phosphoprotein. Eur J Biochem, 1980, 110: 273-278
48. Curtin P, McHugh KP, Zhou H-Y, Fluckiger R, Goldhaber P, Oppenheim FG, Salih E. Modulation of bone resorption by phosphorylation state of bone sialoprotein. Biochemistry, 2009,48:6876-6886
49. Cutting JA, Roth TF. Staining of phospho-proteins on acrylamide gel electropherograms. Anal Biochem, 1973,54:386-394
50. Czernick D, Liu J, Serge D, Salih E. Topographical distribution of phosphorylation sites of phosvitins by mass spectrometry. J Proteomics, 2013,83: 76-98
51. Dahlin S, Angstrom J, Linde A. Dentin phosphoprotein sequence motifs and molecular modeling: conformational adaptations to mineral crystals. Eur J Oral Sci,1998,106:239-248
52. Damodaran S, Xu S. The role of electrostatic forces in anomalous adsorption behavior of phosvitin at the air/water interface. J Colloid Interface Sci, 1996, 178: 426-435
53. Dorozhkin SV. Nanodimensional and nanocrystalline calcium orthophosphates. Biomed. Mater. Diagn. Devices, 2012:19
54. Doucet D, Otter DE, Gauthier SF, Foegeding EA. Enzyme-induced gelation of extensively hydrolyzed whey proteins by Alcalase: peptide identification and determination of enzyme specificity. JAgric Food Chem, 2003,51:6300-6308
55. Dunker AK, Brown CJ, Lawson JD, Iakoucheva LM, Obradovic Z. Intrinsic disorder and protein function. Biochemistry, 2002, 41:6573-6582
56. Feng F, Mine Y. Phosvitin phosphopeptides increase iron uptake in a Caco-2 cell monolayer model. Int J Food Sci Technol, 2006, 41: 455-458
57. Ferraretto A, Gravaghi C, Fiorilli A, Tettamanti G. Casein-derived bioactive phosphopeptides:role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells. FEBS Lett, 2003,551:92-98
58. Finn RN. Vertebrate yolk complexes and the functional implications of phosvitins and other subdomains in vitellogenins. Biol. Reprod,2007,76:926-935
59. Finn RN, Kristoffersen BA. Vertebrate vitellogenin gene duplication in relation to the "3R hypothesis": correlation to the pelagic egg and the oceanic radiation of teleosts. PLoS One, 2007,2:e169
60. Finn RN, Ostby GC, Norberg B, Fyhn HJ. In vivo oocyte hydration in Atlantic halibut (Hippoglossus hippoglossus); proteolytic liberation of free amino acids, and ion transport, are driving forces for osmotic water influx. J Exp Biol, 2002, 205:211-224
61. Fologea D, Ledden B, McNabb DS, Li J. Electrical characterization of protein molecules by a solid-state nanopore. Appl Phys Lett, 2007, 91:053901-053903
62. Francis G, Fisher D, Delgado C, Malik F, Gardiner A, Neale D. PEGylation of cytokines and other therapeutic proteins and peptides: the importance of biological optimisation of coupling techniques. Int J Hematol, 1998, 68:1-18
63. Geladopoulos TP, Sotiroudis TG. Interaction of aluminum ions with phosvitin. J Inorg Biochem,1994,54: 247-256
64. George A, Veis A. Phosphorylated proteins and control over apatite nucleation, crystal growth, and inhibition. Chem Rev, 2008, 108:4670-4693
65. Gericke A, Qin C, Spevak L, Fujimoto Y, Butler W, Sorensen ES, Boskey A. Importance of phosphorylation for osteopontin regulation of biomineralization. Calcif Tissue Int, 2005, 77:45-54
66. Glimcher MJ. Bone:Nature of the calcium phosphate crystals and cellular, structural, and physical chemical mechanisms in their formation. Rev Mineral Geochem,2006,64:223-282
67. Goulas A, Triplett EL, Taborsky G. Oligophosphopeptides of varied structural complexity derived from the egg phosphoprotein, phosvitin. Protein J, 1996, 15: 1-9
68. Greengard O, Sentenac A, Mendelsohn N. Phosvitin, the iron carrier of egg yolk. Biochim Biophys Acta, 1964, 90: 406-407
69. Grizzuti K, Perlmann GE. Conformation of the phosphoprotein, phosvitin. J Biol Chem,1970,245:2573-2578
70. Grizzuti K, Perlmann GE. Binding of magnesium and calcium ions to the phosphoglycoprotein phosvitin. Biochemistry, 1973,12:4399-4403
71. Grizzuti K, Perlmann GE. Further studies on the binding of divalent cations to the phosphoglycoprotein phosvitin. Biochemistry, 1975,14:2171-2175
72. Grogan J, Taborsky G. Iron binding by phosvitin: variation of rate of iron release as a function of the degree of saturation of iron binding sites. J Inorg Biochem, 1986,26:229-236
73. Grohe B, O'Young J, Ionescu DA, Lajoie G, Rogers KA, Karttunen M, Goldberg HA, Hunter GK. Control of calcium oxalate crystal growth by face-specific adsorption of an osteopontin phosphopeptide. J Am Chem Soc, 2007, 129: 14946-14951
74. Guerin-Dubiard C, Anton M, Dhene-Garcia A, Martinet V, Brule G. Hen egg and fish egg phosvitins: composition and iron binding properties. Eur Food Res Technol, 2002, 214:460-464
75. Honig W, Kula M-R. Selectivity of protein precipitation with polyethylene glycol fractions of various molecular weights. Anal Biochem, 1976, 72:502-512
76. He G, Dahl T, Veis A, George A. Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1. Nat Mater, 2003,2:552-558
77. He G, Ramachandran A, Dahl T, George S, Schultz D, Cookson D, Veis A, George A. Phosphorylation of phosphophoryn is crucial for its function as a mediator of biomineralization. JBiol Chem,2005,280:33109-33114
78. Hegenauer J, Saltman P, Nace G. Iron(III)-phosphoprotein chelates: Stoichiometric equilibrium constant for interaction of iron(iii) and phosphorylserine residues of phosvitin and casein. Biochemistry, 1979, 18: 3865-3879
79. Hiramatsu N, Cheek AO, Sullivan CV, Matsubara T, Hara A. Vitellogenesis and endocrine disruption. Biochem Mol Biol Fishes, 2005,6:431-471
80. Hiramatsu N, Ichikawa N, Fukada H, Fujita T, Sullivan CV, Hara A. Identification and characterization of proteases involved in specific proteolysis of vitellogenin and yolk proteins in salmonids. JExp Zool, 2002, 292:11-25
81. Hiramatsu N, Matsubara T, Fujita T, Sullivan CV, Hara A. Multiple piscine vitellogenins:biomarkers of fish exposure to estrogenic endocrine disruptors in aquatic environments. Mar Biol, 2006, 149: 35-47
82. Huang H-D, Lee T-Y, Tzeng S-W, Horng J-T. KinasePhos:a web tool for identifying protein kinase-specific phosphorylation sites. Nucleic Acids Res, 2005, 33:W226-W229
83. Hunter G, Hauschka P, Poole A, Rosenberg L, Goldberg H. Nucleation and inhibition of hydroxyapatite formation by mineralized tissue proteins. Biochem J, 1996,317:59-64
84. Hunter GK, Goldberg HA. Modulation of crystal formation by bone phosphoproteins:role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein. Biochem J, 1994, 302:175-179
85. Huq NL, Cross KJ, Ung M, Reynolds EC. A review of protein structure and gene organisation for proteins associated with mineralised tissue and calcium phosphate stabilisation encoded on human chromosome 4. Arch Oral Biol, 2005, 50: 599-609
86. Ibanez A, Peinado-Onsurbe J, Sanchez E, Prat F. The role of lipoprotein lipase (LPL) in the incorporation of neutral lipids into the oocytes of the European sea bass (Dicentrarchus labrax L.) during gonadal development. Fish Physiol Biochem, 2003,28:291-293
87. Ingham KC. Precipitation of proteins with polyethylene glycol. Method Enzymol, 1989,182:301-306
88. Ishikawa SI, Yano Y, Arihara K, Itoh M. Egg yolk phosvitin inhibits hydroxyl radical formation from the fenton reaction. Biosci, Biotechnol, Biochem, 2004, 68: 1324-1331
89. Ito S, Iijima M, Motai F, Mizoguchi I, Saito T. Effects of calcium salts of acidic monomers on mineral induction of phosphoprotein immobilized to agarose beads. J Biomed Mater Res Part A,2012,100:2760-2765
90. Itoh T, Abe Y, Adachi S. Comparative studies on the alpha and beta-phosvitin from hen's egg yolk. J Food Sci, 1983,48:1755-1757
91. Jiang B, Mine Y. Preparation of novel functional oligophosphopeptides from hen egg yolk phosvitin. JAgric Food Chem, 2000,48:990-994
92. Jiang B, Mine Y. Phosphopeptides derived from hen egg yolk phosvitin: effect of molecular size on the calcium-binding properties. Biosci, Biotechnol, Biochem, 2001,65:1187-1190
93. Joubert FJ, Cook WH. Preparation and characterization of phosvitin from hen egg yolk. Can JBiochem Physiol, 1958, 36:399-408
94. Jung S, Kim DH, Son JH, Nam K, Ahn DU, Jo C. The functional property of egg yolk phosvitin as a melanogenesis inhibitor. Food Chem, 2012, 135:993-998
95. Karampas IA, Kontoyannis CG. Characterization of calcium phosphates mixtures. Vib Spectrosc,2013,64:126-133
96. Kato A, Miyazaki S, Kawamoto A, KOBAYASHI K. Effects of phosphate residues on the excellent emulsifying properties of phosphoglycoprotein phosvitin (food & nutrition). Agric Biol Chem,1987,51:2989-2994
97. Keykhosravani M, Doherty-Kirby A, Zhang C, Brewer D, Goldberg HA, Hunter GK, Lajoie G. Comprehensive identification of post-translational modifications of rat bone osteopontin by mass spectrometry. Biochemistry, 2005,44: 6990-7003
98. Khan MAS, Nakamura S, Ogawa M, Akita E, Azakami H, Kato A. Bactericidal action of egg yolk phosvitin against Escherichia coli under thermal stress. J Agric Food Chem,2000,48:1503-1506
99. Ko KY, Nam KC, Jo C, Lee EJ, Ahn DU. A simple and efficient method for preparing partially purified phosvitin from egg yolk using ethanol and salts. Poultry Sci, 2011,90:1096-1104
100.Kobayashi N, Onuma K, Oyane A, Yamazaki A. The role of phosvitin for nucleation of calcium phosphates on collagen. Key Eng Mater, 2004, 254-256: 537-540
101. Kozlowski H, Mangani S, Messori L, Orioli PL, Scozzafava A. CD and EXAFS study of the interaction between phosvitin and copper(II) ions. J Inorg Biochem, 1988,34: 221-239
102. Kumar V, Sharma VK, Kalonia DS. Effect of polyols on polyethylene glycol (PEG)-induced precipitation of proteins:Impact on solubility, stability and conformation. Int J Pharm,2009,366:38-43
103. Kwak S-Y, Wiedemann-Bidlack FB, Beniash E, Yamakoshi Y, Simmer JP, Litman A, Margolis HC. Role of 20-kDa amelogenin (P148) phosphorylation in calcium phosphate formation in vitro. JBiol Chem, 2009, 284: 18972-18979
104. Lea C, Hawke J. Lipovitellin. 2. The influence of water on the stability of lipovitellin and the effects of freezing and of drying. Biochem J, 1952, 52: 105-114
105. Lee SK, Han JH, Decker EA. Antioxidant activity of phosvitin in phosphatidylcholine liposomes and meat model systems. J Food Sci, 2002, 67: 37-41
106. Lee SL, Veis A, Glonek T. Dentin phosphoprotein: an extracellular calcium-binding protein. Biochemistry, 1977, 16:2971-2979
107. Lei B, Wu J. Purification of egg yolk phosvitin by anion exchange chromatography. JChromatogr A,2012, 1223:41-46
108. Levene P, Alsberg C. Zur Chemie der Paranucleinsaure. Hoppe-Seyler's Zeitschrift fur physiologische Chemie, 1901,31:543-555
109. Li B, Raff J, Barkleit A, Bernhard G, Foerstendorf H. Complexation of U(VI) with highly phosphorylated protein, phosvitin. A vibrational spectroscopic approach. J Inorg Biochem,2010,104: 718-725
110. Lim EH, Teo BY, Lam TJ, Ding JL. Sequence analysis of a fish vitellogenin cDNA with a large phosvitin domain. Gene, 2001,277:175-186
111. Linde A, Lussi A, Crenshaw M. Mineral induction by immobilized polyanionic proteins. Calcif Tissue Int,1989, 44: 286-295
112. Lipmann FA, Levene P. Serinephosphoric acid obtained on hydrolysis of vitellinic acid. JBiol Chem,1932,98:109-114
113. Llena C, Forner L, Baca P. Anticariogenicity of casein phosphopeptide-amorphous calcium phosphate: a review of the literature. J Contemp Dent Pract, 2009, 10:1-9
114.Losso J, Nakai S. A simple procedure for the isolation of phosvitin from chicken egg yolk. In: ed. Egg Uses and Processing Technologies:New Developments. Oxon, UK: Cab International,1994:150-157
115. Lu C-L, Baker RC. Effect of pH and Food Ingredients on the Stability of Egg Yolk Phospholipids and the Metal-Chelator Antioxidant Activity of Phosvitin. J Food Sci,1987,52:613-616
116. Ma J, Wang H, Wang Y, Zhang S. Endotoxin-neutralizing activity of hen egg phosvitin. Mol Immunol,2013,53:355-362
117. Mann S. Mineralization in biological systems. In: ed. Inorganic Elements in Biochemistry: Springer Berlin Heidelberg, 1983:125-174
118. Mann S. Molecular tectonics in biomineralization and biomimetic materials chemistry. Nature, 1993,365:499-505
119. Matsudomi N, Ito K, Yoshika Y. Preventive effect of egg yolk phosvitin on heat-insolubilization of egg white protein and its application to heat-induced egg white gel. Biosci, Biotechnol, Biochem, 2006, 70:836-842
12O. Mecham DK, Olcott HS. Phosvitin, the principal phosphoprotein of egg yolk. J Am Chem Soc,1949,71:3670-3679
121. Meisel H, Meisel H, Fairweather-Tait S, FitzGerald RJ, Hartmann R, Lane C, McDonagh D, Teucher B, Wal J. Detection of caseinophosphopeptides in the distal ileostomy fluid of human subjects. Brit J Nutr, 2003,89:351-358
122. Mikroulis D, Mavrilas D, Kapolos J, Koutsoukos P, Lolas C. Physicochemical and microscopical study of calcific deposits from natural and bioprosthetic heart valves. Comparison and implications for mineralization mechanism. J Mater Sci: Mater Med, 2002, 13:885-889
123. Mine Y. Egg bioscience and biotechnology: John Wiley & Sons, 2008:42-47
124. Moran E. Nutrition of the developing embryo and hatchling. Poultry Sci, 2007, 86: 1043-1049
125. Mosconi G, Carnevali O, Habibi H, Sanyal R, Polzonetti-Magni A. Hormonal mechanisms regulating hepatic vitellogenin synthesis in the gilthead sea bream, Sparus aurata. Am JPhysiol-cell Ph, 2002, 283:C673-C678
126. Nakamura S, Ogawa M, Nakai S, Kato A, Kitts DD. Antioxidant activity of a Maillard-type phosvitin-galactomannan conjugate with emulsifying properties and heat stability. J Agric Food Chem,1998, 46: 3958-3963
127. Obenauer JC, Cantley LC, Yaffe MB. Scansite 2.0: Proteome-wide prediction of cell signaling interactions using short sequence motifs. Nucleic Acids Res, 2003, 31:3635-3641
128. Onuma K. Effect of phosvitin on the nucleation and growth of calcium phosphates in physiological solutions. JPhys Chem B, 2005, 109:8257-8262
129. Osaki S, Sexton RC, Pascual E, Frieden E. Iron oxidation and transferrin formation by phosvitin. Biochem J, 1975, 151:519-525
130. Oshiro M, Yamaguchi K, Takamizawa T, Inage H, Watanabe T, Irokawa A, Ando S, Miyazaki M. Effect of CPP-ACP paste on tooth mineralization: an FE-SEM study. Journal of Oral Science, 2007, 49: 115-120
131. Palmer LC, Newcomb CJ, Kaltz SR, Spoerke ED, Stupp SI. Biomimetic systems for hydroxyapatite mineralization inspired by bone and enamel. Chem Rev, 2008, 108:4754-4783
132. Pan M, Bell WJ, Telfer WH. Vitellogenic blood protein synthesis by insect fat body. Science, 1969, 165:393-394
133. Pasupuleti VK, Braun S. State of the art manufacturing of protein hydrolysates. In: ed. Protein Hydrolysates in Biotechnology: Springer Netherlands, 2010:11-32
134. Perlmann GE, Grizzuti K. Conformational transition of the phosphoprotein phosvitin. Random conformation-β structure. Biochemistry, 1971,10:258-264
135. Prescott B, Renugopalakrishnan V, Glimcher MJ, Bhushan A, Thomas Jr GJ. A Raman spectroscopic study of hen egg yolk phosvitin: structures in solution and in the solid state. Biochemistry,1986,25:2792-2798
136. Raldua D, Fabra M, Bozzo MG, Weber E, Cerda J. Cathepsin B-mediated yolk protein degradation during killifish oocyte maturation is blocked by an H+-ATPase inhibitor: effects on the hydration mechanism. Am J Physiol: Regul, Integr Comp Physiol, 2006, 290: R456-R466
137. Reese C, Heise F, Mayer A. Trans-SNARE pairing can precede a hemifusion intermediate in intracellular membrane fusion. Nature,2005,436:410-414
138. Ren J. Phosvitin extraction and phosphopeptides characterization from chicken egg yolk [Master's thesis]. Edmonton: University of Alberta Libraries, 2012.
139. Renugopalakrishnan V, Horowitz PM, Glimcher MJ. Structural studies of phosvitin in solution and in the solid state. JBiol Chem 1985, 260: 11406-11413
140.Reynolds E. Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions. J Dent Res, 1997, 76: 1587-1595
141. Reynolds EC. Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phosphopeptides: a review. Spec Care Dentist, 1998, 18:8-16
142. Romano M, Rosanova P, Anteo C, Limatola E. Vertebrate yolk proteins: A review. Mol Reprod Dev, 2004, 69:109-116
143. Saad FA, Salih E, Glimcher MJ. Identification of osteopontin phosphorylation sites involved in bone remodeling and inhibition of pathological calcification. J Cell Biochem,2008,103:852-856
144. Saito T, Arsenault AL, Yamauchi M, Kuboki Y, Crenshaw MA. Mineral induction by immobilized phosphoproteins. Bone, 1997, 21:305-311
145. Salih E, Wang J, Mah J, Fluckiger R. Natural variation in the extent of phosphorylation of bone phosphoproteins as a function of in vivo new bone formation induced by demineralized bone matrix in soft tissue and bony environments. Biochem J, 2002, 364:465-474
146. Salih E, Zhou H-Y, Glimcher MJ. Phosphorylation of purified bovine bone sialoprotein and osteopontin by protein kinases. J Biol Chem, 1996, 271: 16897-16905
147. Samaraweera H, Zhang WG, Lee EJ, Ahn DU. Egg yolk phosvitin and functional phosphopeptides-Review. J Food Sci, 2011,76: R143-R150
148. Samaraweera Mudiyanselage CH. Production and characterization of phosphopeptides from egg yolk phosvitin [Ph D dissertation]. Ames:Iowa State University Libraries, 2012.
149. Sappington TW, Raikhel AS. Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem Mol Biol, 1998, 28:277-300
150. Selman K, Wallace RA, Cerda J. Bafilomycin A1 inhibits proteolytic cleavage and hydration but not yolk crystal disassembly or meiosis during maturation of sea bass oocytes. JExp Zool, 2001, 290:265-278
151. Sen LC, Gonzalez-Flores E, Feeney RE, Whitaker JR. Reactions of phosphoproteins in alkaline solutions. JAgric Food Chem, 1977, 25:632-638
152. Shainkin R, Perlmann GE. Phosvitin, a phosphoglycoprotein. I. Isolation and characterization of a glycopeptide from phosvitin. J Biol Chem, 1971a, 246: 2278-2284
153. Shainkin R, Perlmann GE. Phosvitin, a phosphoglycoprotein: Composition and partial structure of carbohydrate moiety. Arch Biochem Biophys, 1971b, 145: 693-700
154. Sharrock WJ, Rosenwasser TA, Gould J, Knott J, Hussey D, Gordon JI, Banaszak L. Sequence of lamprey vitellogenin. Implications for the lipovitellin crystal structure. Journal of Molecular Biology, 1992, 226: 903-907
155. Shkilnyy A, Friedrich A, Tiersch B, Schone S, Fechner M, Koetz J, Schlapfer C-W, Taubert A. Poly (ethylene imine)-controlled calcium phosphate mineralization. Langmuir, 2008,24:2102-2109
156. Shkilnyy A, Schone S, Rumplasch C, Uhlmann A, Hedderich A, Giinter C, Taubert A. Calcium phosphate mineralization with linear poly (ethylene imine): a time-resolved study. Colloid Polym Sci, 2011,289:881-888
157. Silva R, Fischer A, Burch J. The major and minor chicken vitellogenin genes are each adjacent to partially deleted pseudogene copies of the other Mol Cell Biol, 1989,9:3557-3562
158. Silversand C, Haux C. Fatty acid composition of vitellogenin from four teleost species. Journal of Comparative Physiology B, 1995, 164:593-599
159. Sobota M, Salejda A, Trziszka T. Effect of the phosvitin isolation metod on yield of the process. In: 3rd Baltic Conference on Food Science and Technology FOODBALT-2008. Jelgava; 2008:148-151.
160. Spackman DH, Stein WH, Moore S. Automatic recording apparatus for use in chromatography of amino acids. Anal Chem, 1958, 30:1190-1206
161. Sreenath T, Thyagarajan T, Hall B, Longenecker G, D'Souza R, Hong S, Wright JT, MacDougall M, Sauk J, Kulkarni AB. Dentin sialophosphoprotein knockout mouse teeth display widened predentin zone and develop defective dentin mineralization similar to human dentinogenesis imperfecta type III. J Biol Chem, 2003,278:24874-24880
162. Stetler-Stevenson W, Veis A. Bovine dentin phosphophoryn: calcium ion binding properties of a high molecular weight preparation. Calcif Tissue Int, 1987, 40: 97-102
163. Sun C, Hu L, Liu S, Hu G, Zhang S. Antiviral activity of phosvitin from zebrafish Danio rerio. Dev Comp Immunol, 2013, 40:28-34
164. Sundararajan TA, Sampath Kumar KSV, Sarma PS. A simplified procedure for the preparation of phosvitin and vitellin. Biochim Biophys Acta, 1960, 38:360-362
165. Sutton RB, Fasshauer D, Jahn R, Brunger AT. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature, 1998, 395: 347-354
166. Taborsky G. Interaction between phosvitin and iron and its effect on a rearrangement of phosvitin structure. Biochemistry, 1963, 2:266-271
167. Taborsky G. Optical rotatory dispersion and circular dichroism of phosvitin at low pH. Reversible transition between unordered conformation and beta structure. J Biol Chem,1968,243:6014-6020
168. Taborsky G. Effect of freezing and thawing on the conformation of phosvitin. J Biol Chem,1970,245:1054-1062
169. Taborsky G. Iron binding by phosvitin and its conformational consequences. J Biol Chem,1980,255:2976-2984
170. Taborsky G. Phosvitin. Advances in inorganic biochemistry, 1983,5:235-279
171. Taborsky G. On the interaction of phosvitins with ferric ion: Solubility of the Fe(III)-phosphoprotein complex under acidic conditions is a function of the iron/phosphate ratio and the degree of phosvitin phosphorylation. J Inorg Biochem, 1991,44:65-77
172. Taborsky G, Allende CC. A rearrangement in the structure of phosvitin. Biochemistry, 1962,1:406-411
173. Taborsky G, Mok CC. Phosvitin. Homogeneity and molecular weight. J Biol Chem,1967,242:1495-1501
174. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 2011,28:2731-2739
175. Tay F, Pashley D. Biomimetic remineralization of resin-bonded acid-etched dentin. J Dent Res,2009,88:719-724
176. Ternes W. Characterization of water soluble egg yolk proteins with isoelectric focusing. J Food Sci, 1989, 54: 764-765
177. Thompson JR, Banaszak LJ. Lipid-protein interactions in lipo vitellin. Biochemistry,2002,41:9398-9409
178. Veis A. Mineralization in organic matrix frameworks. Reviews in Mineralogy and Geochemistry, 2003, 54: 249-289
179. Vogel HJ. Structure of hen phosvitin: A 31P NMR, 1H NMR, and laser photochemically induced dynamic nuclear polarization 1H NMR study. Biochemistry, 1983,22:668-674
18O. Wallace RA, Morgan JP. Chromatographic resolution of chicken phosvitin. Multiple macromolecular species in a classic vitellogenin-derived phosphoprotein. Biochem J, 1986a, 240:871-878
181. Wallace RA, Morgan JP. Isolation of phosvitin: Retention of small molecular weight species and staining characteristics on electrophoretic gels. Anal Biochem, 1986b,157: 256-261
182. Wallace RA, Selman K. Ultrastructural aspects of oogenesis and oocyte growth in fish and amphibians. J Electron Microsc Tech, 1990, 16:175-201
183. Wallwork M, Kirkham J, Chen H, Chang S, Robinson C, Smith D, Clarkson B. Binding of dentin noncollagenous matrix proteins to biological mineral crystals: an atomic force microscopy study. Calcif Tissue Int, 2002, 71:249-256
184. Wang S-Y, Williams D. Biosynthesis of the vitellogenins. Identification and characterization of nonphosphorylated precursors to avian vitellogenin I and vitellogenin Ⅱ. JBiol Chem, 1982, 257:3837-3846
185. Wang S, Smith DE, Williams DL. Purification of avian vitellogenin III: comparison with vitellogenins I and II. Biochemistry, 1983,22: 6206-6212
186. Wang S, Wang Y, Ma J, Ding Y, Zhang S. Phosvitin plays a critical role in the immunity of zebrafish embryos via acting as a pattern recognition receptor and an antimicrobial effector. JBiol Chem,2011,286:22653-22664
187. Wang SY, Williams DL. Identification, purification, and characterization of two distinct avian vitellogenins. Biochemistry, 1980, 19:1557-1563
188. Wiedemann-Bidlack FB, Kwak S-Y, Beniash E, Yamakoshi Y, Simmer JP, Margolis HC. Effects of phosphorylation on the self-assembly of native full-length porcine amelogenin and its regulation of calcium phosphate formation in vitro. J Struct Biol, 2011,173:250-260
189. Xiao S, Yu C, Chou X, Yuan W, Wang Y, Bu L, Fu G, Qian M, Yang J, Shi Y. Dentinogenesis imperfecta 1 with or without progressive hearing loss is associated with distinct mutations in DSPP. Nat Genet, 2001, 27:201-204
19O. Yamamura JI, Adachi T, Aoki N, Nakajima H, Nakamura R, Matsuda T. Precursor-product relationship between chicken vitellogenin and the yolk proteins: The 40 kDa yolk plasma glycoprotein is derived from the C-terminal cysteine-rich domain of vitellogenin II. Biochim Biophys Acta, 1995, 1244: 384-394
191. Yasui SC, Pancoska P, Dukor RK, Keiderling TA, Renugopalakrishnan V, Glimcher MJ, Clark RC. Conformational transitions in phosvitin with pH variation. Vibrational circular dichroism study. JBiol Chem, 1990, 265:3780-3788
192. Yoshizaki N, Yonezawa S. Cathepsin D activity in the vitellogenesis of Xenopus laevis. Dev, Growth Differ, 1994,36:299-306
193. Young D, Nau F, Pasco M, Mine Y. Identification of hen egg yolk-derived phosvitin phosphopeptides and their effects on gene expression profiling against oxidative stress-induced caco-2 cells. JAgric Food Chem, 2011,59:9207-9218
194. Zachariou M, Traverso I, Spiccia L, Hearn MT. Potentiometric investigations into the acid-base and metal ion binding properties of immobilized metal ion affinity chromatographic (IMAC) adsorbents. JPhys Chem, 1996, 100:12680-12690
195. Zhang X, Qiu N, Geng F, Ma M. Simply and effectively preparing high-purity phosvitin using polyethylene glycol and anion-exchange chromatography. J Sep Sci,2011,34:3295-3301
196. Znidarsic WJ, Chen IW, Shastri VP. Influence of surface charge and protein intermediary layer on the formation of biomimetic calcium phosphate on silica nanoparticles. J Mater Chem, 2012, 22:19562-19569
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |