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Chemistry for the analysis of protein–protein interactions: Rapid and efficient cross-linking triggered by long wavelength light

By David A. Fancy and Thomas Kodadek

Abstract

Chemical cross-linking is a potentially useful technique for probing the architecture of multiprotein complexes. However, analyses using typical bifunctional cross-linkers often suffer from poor yields, and large-scale modification of nucleophilic side chains can result in artifactual results attributable to structural destabilization. We report here the de novo design and development of a type of protein cross-linking reaction that uses a photogenerated oxidant to mediate rapid and efficient cross-linking of associated proteins. The process involves brief photolysis of tris-bipyridylruthenium(II) dication with visible light in the presence of the electron acceptor ammonium persulfate and the proteins of interest. Very high yields of cross-linked products can be obtained with irradiation times of <1 second. This chemistry obviates many of the problems associated with standard cross-linking reagents

Topics: Physical Sciences
Publisher: The National Academy of Sciences
Year: 1999
OAI identifier: oai:pubmedcentral.nih.gov:26828
Provided by: PubMed Central
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