The SH3 domain-binding T cell tyrosyl phosphoprotein p120. Demonstration of its identity with the c-cbl protooncogene product and in vivo complexes with Fyn, Grb2, and phosphatidylinositol 3-kinase

T. Fukazawa, K. A. Reedquist, T. Trub, S. Soltoff, G. Panchamoorthy, B. Druker, L. Cantley, S. E. Shoelson, Hamid Band

Research output: Contribution to journalArticle

184 Citations (Scopus)

Abstract

Previously, we have identified p120 as a Fyn/Lck SH3 and SH2 domain- binding protein that is tyrosine phosphorylated rapidly after T cell receptor triggering. Here, we used direct protein purification, amino acid sequence analysis, reactivity with antibodies, and two-dimensional gel analyses to identify p120 as the human c-cbl protooncogene product. We demonstrate in vivo complexes of p120(cbl) with Fyn tyrosine kinase, the adaptor protein Grb2, and the p85 subunit of phosphatidylinositol (PI) 3-kinase. The association of p120(cbl) with Fyn and the p85 subunit of PI 3-kinase (together with PI 3-kinase activity) was markedly increased by T cell activation, consistent with in vitro binding of p120(cbl) to their SH2 as well as SH3 domains. In contrast, a large fraction of p120(cbl) was associated with Grb2 prior to activation, and this association did not change upon T cell activation. In vitro, p120(cbl) interacted with Grb2 exclusively through its SH3 domains. These results demonstrate a novel Grb2-p12(cbl) signaling complex in T cells, distinct from the previously analyzed Grb2-Sos complex. The association of p120(cbl) with ubiquitous signaling proteins strongly suggests a general signal transducing function for this enigmatic protooncogene with established leukemogenic potential but unknown physiological function.

Original languageEnglish (US)
Pages (from-to)19141-19150
Number of pages10
JournalJournal of Biological Chemistry
Volume270
Issue number32
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

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Oncogene Protein v-cbl
Phosphatidylinositol 3-Kinase
src Homology Domains
T-cells
Phosphoproteins
Demonstrations
T-Lymphocytes
Chemical activation
Association reactions
Proto-Oncogene Proteins c-fyn
Proteins
Protein Sequence Analysis
T-Cell Antigen Receptor
Purification
Tyrosine
Carrier Proteins
Gels
Amino Acids
Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The SH3 domain-binding T cell tyrosyl phosphoprotein p120. Demonstration of its identity with the c-cbl protooncogene product and in vivo complexes with Fyn, Grb2, and phosphatidylinositol 3-kinase. / Fukazawa, T.; Reedquist, K. A.; Trub, T.; Soltoff, S.; Panchamoorthy, G.; Druker, B.; Cantley, L.; Shoelson, S. E.; Band, Hamid.

In: Journal of Biological Chemistry, Vol. 270, No. 32, 01.01.1995, p. 19141-19150.

Research output: Contribution to journalArticle

Fukazawa, T. ; Reedquist, K. A. ; Trub, T. ; Soltoff, S. ; Panchamoorthy, G. ; Druker, B. ; Cantley, L. ; Shoelson, S. E. ; Band, Hamid. / The SH3 domain-binding T cell tyrosyl phosphoprotein p120. Demonstration of its identity with the c-cbl protooncogene product and in vivo complexes with Fyn, Grb2, and phosphatidylinositol 3-kinase. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 32. pp. 19141-19150.
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abstract = "Previously, we have identified p120 as a Fyn/Lck SH3 and SH2 domain- binding protein that is tyrosine phosphorylated rapidly after T cell receptor triggering. Here, we used direct protein purification, amino acid sequence analysis, reactivity with antibodies, and two-dimensional gel analyses to identify p120 as the human c-cbl protooncogene product. We demonstrate in vivo complexes of p120(cbl) with Fyn tyrosine kinase, the adaptor protein Grb2, and the p85 subunit of phosphatidylinositol (PI) 3-kinase. The association of p120(cbl) with Fyn and the p85 subunit of PI 3-kinase (together with PI 3-kinase activity) was markedly increased by T cell activation, consistent with in vitro binding of p120(cbl) to their SH2 as well as SH3 domains. In contrast, a large fraction of p120(cbl) was associated with Grb2 prior to activation, and this association did not change upon T cell activation. In vitro, p120(cbl) interacted with Grb2 exclusively through its SH3 domains. These results demonstrate a novel Grb2-p12(cbl) signaling complex in T cells, distinct from the previously analyzed Grb2-Sos complex. The association of p120(cbl) with ubiquitous signaling proteins strongly suggests a general signal transducing function for this enigmatic protooncogene with established leukemogenic potential but unknown physiological function.",
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AU - Fukazawa, T.

AU - Reedquist, K. A.

AU - Trub, T.

AU - Soltoff, S.

AU - Panchamoorthy, G.

AU - Druker, B.

AU - Cantley, L.

AU - Shoelson, S. E.

AU - Band, Hamid

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