Selective binding of activated pp60c-src by an immobilized synthetic phosphopeptide modeled on the carboxyl terminus of pp60c-src.
Open Access
- 1 December 1991
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 88 (23), 10696-10700
- https://doi.org/10.1073/pnas.88.23.10696
Abstract
Phosphorylation of the carboxyl terminus of pp60c-src, the product of the c-src protooncogene, at Tyr-527 suppresses its tyrosine kinase activity and transforming potential. It has been proposed that the phosphorylated carboxyl terminus of pp60c-src inhibits kinase activity by binding to the SH2 (src homology 2) domain. We have synthesized peptides corresponding to the carboxyl-terminal 13 residues of pp60c-src phosphorylated and nonphosphorylated at Tyr-527. A highly transforming mutant, pp60c-src(F527), in which Tyr-527 is mutated to Phe, bound to the phosphorylated peptide immobilized to Affi-Gel 10. Binding of the phosphorylated peptide was abolished by deletion of residues 144-175 in the SH2 domain but not by deletion of residues 93-143, which removes most of the SH3 domain. The phosphorylated peptide also bound to pp60v-src, the transforming protein of Rous sarcoma virus. Only traces of pp60v-src and pp60c-src(F527) bound to the corresponding nonphosphorylated c-src peptide. Normal pp60c-src bound much less efficiently to the phosphorylated peptide than did pp60c-src(F527). A phosphorylated peptide corresponding to the carboxyl terminus of the c-fgr protein also bound to pp60c-src(F527), but with weaker affinity. Furthermore, the phosphorylated synthetic carboxyl-terminal pp60c-src peptide markedly inhibited phosphorylation of pp60c-src(F527) during cytoskeletal kinase assays. These results provide direct evidence for models in which the phosphorylated carboxyl terminus of pp60c-src binds intramolecularly or intermolecularly to the SH2 domain of the c-src protein.Keywords
This publication has 35 references indexed in Scilit:
- SH2 and SH3 Domains: Elements that Control Interactions of Cytoplasmic Signaling ProteinsScience, 1991
- Deletions in the SH2 domain of p60v-src prevent association with the detergent-insoluble cellular matrix.Molecular and Cellular Biology, 1991
- Identification of domains of the v-crk oncogene product sufficient for association with phosphotyrosine-containing proteins.Molecular and Cellular Biology, 1991
- The noncatalytic src homology region 2 segment of abl tyrosine kinase binds to tyrosine-phosphorylated cellular proteins with high affinity.Proceedings of the National Academy of Sciences, 1991
- SH2 mutants of c-src that are host dependent for transformation are trans-dominant inhibitors of mouse cell transformation by activated c-src.Genes & Development, 1990
- Src homology region 2 domains direct protein-protein interactions in signal transduction.Proceedings of the National Academy of Sciences, 1990
- Mutations in src homology regions 2 and 3 of activated chicken c-src that result in preferential transformation of mouse or chicken cells.Proceedings of the National Academy of Sciences, 1990
- INVIVO PHOSPHORYLATION AND MEMBRANE ASSOCIATION OF THE FYN PROTOONCOGENE PRODUCT IN IM-9 HUMAN LYMPHOBLASTS1990
- Neoplastic transformation induced by an activated lymphocyte-specific protein tyrosine kinase (pp56lck).Molecular and Cellular Biology, 1988
- Association of p60src with Triton X-100-resistant cellular structure correlates with morphological transformation.Proceedings of the National Academy of Sciences, 1987