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To investigate the nature of proteins recognized by Src homology 3 (SH3) domains, a cDNA expression library was prepared from macrophages and screened with a probe representing the three SH3 domains of p47nck. Two clones were isolated, and one, designated SAKAP I (for Src A box Nck-associated protein I), contained the carboxyl-terminal half of the cbl proto-oncogene product. Studies in vitro demonstrated reactivity between SAKAP I and SH3 domains derived from a variety of molecules. Wide variations in this assay suggested a high degree of specificity inherent in SAKAP I binding. Moreover, it was possible to demonstrate an in vivo association between p47nck and p120c-cbl in HL60 cells. These findings suggest that proteins containing SH3 elements regulate Cbl function.
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