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Quantitative structure-activity analysis correlating Ras/Raf interaction in vitro to Raf activation in vivo

Nature Structural Biology volume 3, pages 244–251 (1996)Cite this article

Abstract

Binding of Ras to c-Raf-1 is a pivotal step of many mitogenic signalling pathways. Based on the recent crystal structure of the complex of Rap1 A with the Ras-binding domain of Raf, mutations were introduced in c-Raf-1 and their effects on Ras/Raf binding affinity in vitro and Ras/Raf regulated gene expression in vivo were analysed. Our data reveal an empirical semi-logarithmic correlation between dissociation constants and Raf-induced gene activity. The functional epitope that primarily determines binding affinity consists of residues Gin 66, Lys 84 and Arg 89 in Raf. This quantitative structure-activity investigation may provide a general approach to correlate structure-guided biochemical analysis with biological function of protein–protein interactions.

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Authors and Affiliations

  1. Max-Planck-lnstitut für Molekulare Physiologie, Abteilung Strukturelle Biologie, Rheinlanddamm 201, D-44139, Dortmund, Federal Republic of Germany

    Christoph Block, Christian Herrmann, Nicolas Nassar & Alfred Wittinghofer

  2. Medizinische Hochschule Hannover, Institut für Molekularbiologie, D-30623, Hannover, Federal Republic of Germany

    Ralf Janknecht

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  1. Christoph Block
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  5. Alfred Wittinghofer
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Block, C., Janknecht, R., Herrmann, C. et al. Quantitative structure-activity analysis correlating Ras/Raf interaction in vitro to Raf activation in vivo. Nat Struct Mol Biol 3, 244–251 (1996). https://doi.org/10.1038/nsb0396-244

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