一种用于探索蛋白质相互作用的模块化交联方法。

A modular cross-linking approach for exploring protein interactions.

作者信息

Trester-Zedlitz Michelle, Kamada Katsuhiko, Burley Stephen K, Fenyö David, Chait Brian T, Muir Tom W

机构信息

Laboratories of Synthetic Protein Chemistry, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.

出版信息

J Am Chem Soc. 2003 Mar 5;125(9):2416-25. doi: 10.1021/ja026917a.

DOI:10.1021/ja026917a

PMID:12603129

Abstract

A method is described for the elucidation of protein-protein interactions using novel cross-linking reagents and mass spectrometry. The method incorporates (1) a modular solid-phase synthetic strategy for generating the cross-linking reagents, (2) enrichment and digestion of cross-linked proteins using microconcentrators, (3) mass spectrometric analysis of cross-linked peptides, and (4) comprehensive computational analysis of the cross-linking data. This integrated approach has been applied to the study of cross-linking between the components of the heterodimeric protein complex negative cofactor 2.

摘要

本文描述了一种使用新型交联试剂和质谱法阐明蛋白质-蛋白质相互作用的方法。该方法包括：(1) 用于生成交联试剂的模块化固相合成策略；(2) 使用微浓缩器对交联蛋白进行富集和消化；(3) 对交联肽进行质谱分析；以及 (4) 对交联数据进行全面的计算分析。这种综合方法已应用于异二聚体蛋白复合物负辅因子2各组分之间交联的研究。