不同吸血时段蜱粪便蛋白组学比较分析
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摘要
为了解蜱血液消化特征,通过耳袋饲养法分别收集镰形扇头蜱雌性成蜱吸血d4、d8的粪便。提取粪便蛋白质,进行SDS-PAGE和LC/MS-MS分析。用非标定量分析蛋白组学技术对蜱不同吸血时段粪便进行差异蛋白质表达谱分析。结果共鉴定蜱源肽段数628个、蛋白质总数196个;鉴定兔源肽段数6173个、蛋白质总数659个。对组间差异蛋白分析发现,除一些未知蛋白外,其中蜱源蛋白主要是热休克蛋白、免疫蛋白相关亚基和一些蛋白酶类等。兔源差异蛋白主要为一些凝血因子、糖蛋白,以及一些物质和能量的消化与合成的相关酶类,如兔丝氨酸/苏氨酸蛋白磷酸酶、三磷酸肌苷焦磷酸酶、S-(羟甲基)谷胱甘肽脱氢酶等。本研究中的粪便蛋白质组反应了蜱血液消化过程的复杂性,丰富了蜱消化方面的研究数据,同时为抗蜱疫苗研究提供了一个思路。
To understand the characteristics of blood digestion, the feces of Rhipicephalus haemaphysaloides female ticks were collected at the fourth and eighth days of sucking blood for protein extraction. The extracted proteins were used for SDS-PAGE analysis and LC/MS-MS. Differentially expressed proteins during different blood-sucking periods were analyzed by label free proteomics techniques. A total of 628 peptides and 196 proteins were identified as tick-derived while 6,173 peptides and 659 proteins were rabbit-derived. Analysis of differential proteins between groups revealed that the tick-derived proteins were mainly heat shock proteins, immunoprotein-related subunits and some proteases besides some unknown proteins while rabbit-derived proteins were mainly coagulation factors, glycoproteins and some related enzymes that involved in some substances, energy digestion and synthesis such as rabbit serine/threonine protein phosphatase, inosine triphosphate pyrophosphatase, S-(hydroxymethyl)glutathione dehydrogenase and so on. The fecal proteome reflected the complexity of tick blood digestion process and needed further investigation.
To understand the characteristics of blood digestion, the feces of Rhipicephalus haemaphysaloides female ticks were collected at the fourth and eighth days of sucking blood for protein extraction. The extracted proteins were used for SDS-PAGE analysis and LC/MS-MS. Differentially expressed proteins during different blood-sucking periods were analyzed by label free proteomics techniques. A total of 628 peptides and 196 proteins were identified as tick-derived while 6,173 peptides and 659 proteins were rabbit-derived. Analysis of differential proteins between groups revealed that the tick-derived proteins were mainly heat shock proteins, immunoprotein-related subunits and some proteases besides some unknown proteins while rabbit-derived proteins were mainly coagulation factors, glycoproteins and some related enzymes that involved in some substances, energy digestion and synthesis such as rabbit serine/threonine protein phosphatase, inosine triphosphate pyrophosphatase, S-(hydroxymethyl)glutathione dehydrogenase and so on. The fecal proteome reflected the complexity of tick blood digestion process and needed further investigation.
引文
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[2]Rodríguez-Mallon A.Developing anti-tick vaccines[J].Methods Mol Biol,2016,1404:243-259.
[3]Valle M R,Guerrero F D.Anti-tick vaccines in the omics era[J].Front Biosci(Elite Ed),2018,10:122-136.
[4]H o r n M,N u s s b a u m e r o váM,S a n d a M,e t a l.Hemoglobin digestion in blood-feeding ticks:mapping a multipeptidase pathway by functional proteomics[J].Chem Biol,2009,16(10):1053-1063.
[5]Mans B J,Neitz A W.Adaptation of ticks to a bloodfeeding environment:evolution from a functional perspective[J].Insect Biochem Mol Biol,2004,34(1):1-17.
[6]Cox J,Mann M.MaxQuant enables high peptide identification rates,individualized p.p.b.-range mass accuracies and proteome-wide protein quantification[J].Nat Biotechnol,2008,26(12):1367-1372.
[7]Calvo E,Mizurini D M,Sá-Nunes A,et al.Alboserpin,a factor Xa inhibitor from the mosquito vector of yellow fever,binds heparin and membrane phospholipids and exhibits antithrombotic activity[J].J Biol Chem,2011,286(32):27998-28010.
[8]Neelakanta G,Sultana H,Sonenshine D E,et al.Identification and characterization of a histamine-binding lipocalin-like molecule from the relapsing fever tick Ornithodoros turicata[J].Insect Mol Biol,2018,27(2):177-187.
[9]Osborne C,West E,Bate C.The phospholipase A2pathway controls a synaptic cholesterol ester cycle and synapse damage[J].J Cell Sci,2018,131(8).pii:jcs211789.
[10]Maya-Monteiro C M,Daffre S,Logullo C,et al.HeLp,a heme lipoprotein from the hemolymph of the cattle tick,Boophilus microplus[J].J Biol Chem,2000,275(47):36584-36589.
[11]Lara F A,Pohl P C,Gandara A C,et al.ATP binding cassette transporter mediates both heme and pesticide detoxification in tick midgut cells[J].PloS One,2015,10(8):e0134779.
[12]Shinde S R,Maddika S.Post translational modifications of Rab GTPases[J].Small GTPases,2018,9(1-2):49-56.
[13]Perner J,Provaznik J,SchrenkováJ,et al.RNA-seq analyses of the midgut from blood-and serum-fed Ixodes ricinus ticks[J].Sci Rep,2016,6:36695.