CROSSLINKING REAGENTS, METHODS, AND COMPOSITIONS FOR STUDYING PROTEIN-PROTEIN INTERACTIONS
申请人:Huang Yumei
公开号:US20120322978A1
公开(公告)日:2012-12-20
The invention provides reagents, methods, and compositions for studying protein-protein interactions. The inventive system and methods allow the analysis of protein-protein interactions in vivo and in vitro. Advantages offered by various embodiments of the inventive system and methods compared to existing photocrosslinking approaches include, for example, (i) novel reversible crosslinking reagents that allow easy isolation, purification, and enrichment of the crosslinked products; (ii) trifluoromethyl phenyldiazirine- or perfluorinated phenylazide-based photocrosslinking reagents that provide high specific labeling, no side product, and higher photocrosslinking efficiency; (iii) versatile spacer groups that allow systematic contact site mapping; (iv) novel methods for isolating, purifying, and detecting crosslinked products based on the reversible-link chemistry; and (v) the ability to study the interaction sites in vitro, in situ, or in vivo.
该发明提供了用于研究蛋白质-蛋白质相互作用的试剂、方法和组合物。这一创新系统和方法允许在体内和体外分析蛋白质-蛋白质相互作用。与现有的光交联方法相比,该创新系统和方法的各种实施例提供的优势包括:(i)新型可逆交联试剂,可方便地分离、纯化和富集交联产物;(ii)三氟甲基苯基重氮基或全氟苯基偶氮基光交联试剂提供高特异标记、无副产物和更高的光交联效率;(iii)多功能的间隔基团,可进行系统的接触位点映射;(iv)基于可逆链接化学的新方法用于分离、纯化和检测交联产物;以及(v)能够在体外、原位或体内研究相互作用位点。