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, 325 (1), 183-90

Mutational Analysis of the Interaction Between Insulin Receptor and IGF-I Receptor With c-Crk and Crk-L in a Yeast Two-Hybrid System

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Mutational Analysis of the Interaction Between Insulin Receptor and IGF-I Receptor With c-Crk and Crk-L in a Yeast Two-Hybrid System

Jürgen Klammt et al. Biochem Biophys Res Commun.

Abstract

The SH2/SH3 adapter proteins of the Crk family are potent signal transducers after receptor tyrosine kinase stimulation with insulin or IGF-1. We have employed a yeast two-hybrid approach and mutational analysis to dissect the capabilities of the insulin receptor and the IGF-I receptor to directly associate with Crk isoforms. Insulin receptor stably recruits full length Crk by association with its SH2 domain in an auto-phosphorylation dependent manner. In contrast, interaction of the IGF-I receptor with the Crk-IISH2 domain was only detectable when Crk-II was truncated in its C-terminal part, indicating the transient nature of this interaction. From these data it can be concluded that members of the insulin receptor family activate Crk proteins in a differential manner.

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