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Src homology domains of v-Src stabilize an active conformation of the tyrosine kinase catalytic domain

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Abstract

To examine the interactions between Src homology,domains and the tyrosine kinase catalytic domain of v-Src, various combinations of domains have been expressed in bacteria as fusion proteins. Constructs containing the isolated catalytic domain, SH2 + catalytic domain, and SH3 + SH2 + catalytic domains were active in autophosphorylation assays. For the catalytic domain of v-Src, but not for v-Abl, addition of exogenous Src SH3-SH2 domains stimulated the autophosphorylation activity. In contrast to results for autophosphorylation, constructs containing Src homology domains were more active towards a synthetic peptide substrate than the isolated catalytic domain. The ability of the SH2 and SH3 domains of v-Src to stabilize an active enzyme conformation was also confirmed by refolding after denaturation in guanidinium hydrochloride. Collectively the data suggest that, in addition to their roles in intermolecular protein-protein interactions, the Src homology regions of v-Src exert a positive influence on tyrosine kinase function, potentially by maintaining an active conformation of the catalytic domain.

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Authors and Affiliations

  1. Department of Physiology and Biophysics, School of Medicine, State University of New York at Stony Brook, 11794, Stony Brook, NY, USA

    Bin Xu & W. Todd Miller

  2. Department of Physiology and Biophysics, Basic Science Tower T-6, School of Medicine, SUNY at Stony Brook, 11794-8661, Stony Brook, NY, USA

    W. Todd Miller

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  1. Bin Xu
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Xu, B., Miller, W.T. Src homology domains of v-Src stabilize an active conformation of the tyrosine kinase catalytic domain. Mol Cell Biochem 158, 57–63 (1996). https://doi.org/10.1007/BF00225883

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