Tyrosine phosphorylation of a common 57-kDa protein in growth factor-stimulated and -transformed cells

J Biol Chem. 1991 Apr 15;266(11):6808-14.

Abstract

Protein tyrosine phosphorylation was studied in macrophages and fibroblasts to identify putative components of post-receptor mitogenic pathways that might be functionally conserved in different cell types. Nondenaturing conditions were established for the approximately quantitative recovery of anti-phosphotyrosine antibody (alpha PY)-reactive proteins from cells. A common, 57-kDa alpha PY-reactive protein was identified by V8 protease peptide mapping in colony-stimulating factor-1 (CSF-1)- or granulocyte-macrophage colony-stimulating factor (GM-CSF)-stimulated BAC1.2F5 macrophages, in platelet-derived growth factor-stimulated NIH-3T3 cells, and in CSF-1-stimulated NIH-3T3 cells expressing the c-fms/CSF-1 receptor. The 57-kDa protein was phosphorylated on serine and tyrosine and was the only alpha PY-reactive protein band whose phosphorylation was reproducibly increased in GM-CSF-stimulated cells. The effect of the growth factors on the tyrosine phosphorylation of the 57-kDa protein could be mimicked by treatment of the cells with orthovanadate, a phosphotyrosine protein phosphatase inhibitor. In the absence of growth factors, tyrosine phosphorylation of the 57-kDa protein was higher in v-fms or c-fms (F969, S301)-transformed NIH-3T3 cells than in untransformed NIH-3T3 (c-fms) and NIH-3T3 (c-fms, F969) cells. These data indicate that the 57-kDa protein is a common target for growth factor-stimulated tyrosine phosphorylation and potentially important for growth factor mitogenic signaling.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Cell Transformation, Neoplastic*
  • Electrophoresis, Gel, Two-Dimensional
  • Electrophoresis, Polyacrylamide Gel
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
  • Growth Substances / pharmacology*
  • Kinetics
  • Macrophage Colony-Stimulating Factor / pharmacology*
  • Mice
  • Molecular Weight
  • Phosphates / metabolism
  • Phosphoproteins / isolation & purification*
  • Phosphorylation
  • Phosphotyrosine
  • Recombinant Proteins / pharmacology
  • Tyrosine* / analogs & derivatives
  • Tyrosine* / analysis

Substances

  • Growth Substances
  • Phosphates
  • Phosphoproteins
  • Recombinant Proteins
  • Phosphotyrosine
  • Tyrosine
  • Macrophage Colony-Stimulating Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor