In Vivo Application of Photocleavable Protein Interaction Reporter Technology
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- 作者：Li Yang ; Chunxiang Zheng ; Chad R. Weisbrod ; Xiaoting Tang ; Gerhard R. Munske ; Michael R. Hoopmann ; Jimmy K. Eng ; James E. Bruce
- 刊名：Journal of Proteome Research
- 出版年：2012
- 出版时间：February 3, 2012
- 年：2012
- 卷：11
- 期：2
- 页码：1027-1041
- 全文大小：796K
- 年卷期：v.11,no.2(February 3, 2012)
- ISSN：1535-3907
文摘
In vivo protein structures and protein鈥損rotein interactions are critical to the function of proteins in biological systems. As a complementary approach to traditional protein interaction identification methods, cross-linking strategies are beginning to provide additional data on protein and protein complex topological features. Previously, photocleavable protein interaction reporter (pcPIR) technology was demonstrated by cross-linking pure proteins and protein complexes and the use of ultraviolet light to cleave or release cross-linked peptides to enable identification. In the present report, the pcPIR strategy is applied to Escherichia coli cells, and in vivo protein interactions and topologies are measured. More than 1600 labeled peptides from E. coli were identified, indicating that many protein sites react with pcPIR in vivo. From those labeled sites, 53 in vivo intercross-linked peptide pairs were identified and manually validated. Approximately half of the interactions have been reported using other techniques, although detailed structures exist for very few. Three proteins or protein complexes with detailed crystallography structures are compared to the cross-linking results obtained from in vivo application of pcPIR technology.