Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family

Paul R Langlais, Lily Q. Dong, Derong Hu, Feng Liu

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Treatment of cells with insulin and protein tyrosine phosphatase inhibitors such as vanadate and pervanadate resulted in the tyrosine phosphorylation of Grb10, a Src homology 2 (SH2) and pleckstrin homology domain-containing adaptor protein which binds to a number of receptor tyrosine kinases including the insulin receptor (IR). Although Grb10 binds directly to the kinase domain of the IR, our data show that Grb10 is not a direct substrate for the IR tyrosine kinase. Consistent with this finding, Grb10 tyrosine phosphorylation in cells was inhibited by herbimycin A, a relatively specific inhibitor for members of the Src tyrosine kinase family, and by the expression of dominant negative Src or Fyn. In addition, Grb10 tyrosine phosphorylation was stimulated by expression of constitutively active Src or Fyn in cells and by incubation with purified Src or Fyn in vitro. The insulin stimulated or Src/Fyn-mediated tyrosine phosphorylation in vivo was significantly reduced when Grb10 tyrosine 67 was changed to glycine. This mutant form of Grb10 bound with higher affinity to the IR in cells than that of the wild-type protein, suggesting that tyrosine phosphorylation of Grb10 may normally negatively regulate its binding to the IR. Our data show that Grb10 is a new substrate for members of the Src tyrosine kinase family and that the tyrosine phosphorylation of the protein may play a potential role in cell signaling processes mediated by these kinases.

Original languageEnglish (US)
Pages (from-to)2895-2903
Number of pages9
JournalOncogene
Volume19
Issue number25
StatePublished - Jun 8 2000
Externally publishedYes

Fingerprint

src-Family Kinases
Tyrosine
Insulin Receptor
Phosphorylation
Phosphotransferases
Insulin
Proteins
Protein Tyrosine Phosphatases
Vanadates
Receptor Protein-Tyrosine Kinases
Glycine

Keywords

  • Grb10
  • IR
  • SH2
  • Src

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Langlais, P. R., Dong, L. Q., Hu, D., & Liu, F. (2000). Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family. Oncogene, 19(25), 2895-2903.

Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family. / Langlais, Paul R; Dong, Lily Q.; Hu, Derong; Liu, Feng.

In: Oncogene, Vol. 19, No. 25, 08.06.2000, p. 2895-2903.

Research output: Contribution to journalArticle

Langlais, PR, Dong, LQ, Hu, D & Liu, F 2000, 'Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family', Oncogene, vol. 19, no. 25, pp. 2895-2903.
Langlais PR, Dong LQ, Hu D, Liu F. Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family. Oncogene. 2000 Jun 8;19(25):2895-2903.
Langlais, Paul R ; Dong, Lily Q. ; Hu, Derong ; Liu, Feng. / Identification of Grb10 as a direct substrate for members of the Src tyrosine kinase family. In: Oncogene. 2000 ; Vol. 19, No. 25. pp. 2895-2903.
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