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Proc Natl Acad Sci U S A. 1999 May 25;96(11):6020-4.

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

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  • 1Departments of Internal Medicine and Biochemistry, Center for Biomedical Inventions, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235-8573, USA.

Erratum in

  • Proc Natl Acad Sci U S A 2000 Feb 1;97(3):1317.


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.

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