SRC HOMOLOGY-2 DOMAINS OF PROTEIN-TYROSINE-PHOSPHATASE ARE PHOSPHORYLATED BY INSULIN-RECEPTOR KINASE AND BIND TO THE COOH-TERMINUS OF INSULIN-RECEPTORS IN-VITRO

To clarify the role of protein tyrosine phosphatase (PTPas) containing Src homology 2 (SH2) regions on insulin signaling, we investigated the interactions between SH2 regions of PTPase and insulin receptors. We made a pair of SH2 domains of PTP1C and SH-PTP2 fusion proteins coupled to glutathione-S-transferase (GST) using pGEX-3X expression vector. After incubating with insulin, insulin receptors were incubated with SH2 proteins in the presence of 100 μ ATP at 4°C for 3 hr, and then immunoprecipitated and analyzed by SDS-PAGE. We found that SH2 domains of SH-PTP2 were phosphorylated, but not those of PTP1C by insulin receptor kinase and the SH2 domains of SH-PTP2, but not those of PTP1C, directly bound to the phosphorylated COOH-terminus of insulin receptos in vitro. © 1993 Academic Press, Inc.