Characterization of a novel tyrosine phosphorylated 100-kDa protein that binds to SHP-2 and phosphatidylinositol 3'-kinase in myeloid cells.

Fms is a tyrosine kinase-containing receptor for macrophage colony-stimulating factor (M-CSF) that regulates survival, growth, and differentiation of cells along the monocyte/macrophage lineage. M-CSF stimulation of murine myeloid FDC-P1 cells expressing Fms resulted in the tyrosine phosphorylation of a number of signal transduction proteins, including an unidentified 100-kDa protein. This 100-kDa protein associated with the tyrosine phosphatase SHP-2 but not with the related phosphatase SHP-1. The kinetics of tyrosine phosphorylation of p100 and SHP-2 suggest that p100 may be a direct substrate of SHP-2. p100 bound directly to the SH2 domains of both SHP-2 and the p85 subunit of phosphatidylinositol 3'-kinase. The 100-kDa protein did not appear to bind directly to Fms, Ship, Cbl, Shc, or Grb2, although all of these proteins were coimmunoprecipitated with p85 after M-CSF stimulation. Association of p100 with SHP-2 and p85 did not require the major autophosphorylation sites on Fms nor binding of p85 to Fms. A tyrosine phosphorylated protein of 100 kDa also coprecipitated with SHP-2 from several other myeloid cell lines after M-CSF stimulation but was not seen in immunoprecipitates from Rat2 fibroblasts expressing Fms. Stimulation of FDC-P1 cells with additional cytokines also resulted in coprecipitation of a 100-kDa protein with SHP-2. p100 may therefore be a common component of the signaling pathways of cytokine receptors in myeloid cells.