Yu J C, Heidaran M A, Pierce J H, Gutkind J S, Lombardi D, Ruggiero M, Aaronson S A
Laboratory of Cellular and Molecular Biology, National Cancer Institute (37-1E24), Bethesda, Maryland 20892.
Mol Cell Biol. 1991 Jul;11(7):3780-5. doi: 10.1128/mcb.11.7.3780-3785.1991.
A phosphatidylinositol-3 (PI-3) kinase activity of unknown biological function associates with tyrosine kinase-containing proteins, including a number of growth factor receptors after ligand stimulation. In the beta platelet-derived growth factor (beta PDGF) receptor, phosphorylation of a specific tyrosine residue within the kinase insert domain was required for its interaction with this enzyme. We show that substitutions of phenylalanine for tyrosine residue 731 or 742 within the kinase insert domain of the alpha PDGF receptor do not impair PDGF-induced tyrosine phosphorylation of the receptor or of an in vivo substrate, phospholipase C-gamma. Moreover, phosphatidylinositol turnover in response to ligand stimulation is unaffected. However, both lesions markedly impair receptor association with PI-3 kinase. Antiphosphotyrosine antibody-recoverable PI-3 kinase was also dramatically reduced in PDGF-stimulated cells expressing either mutant receptor. Since neither mutation abolished PDGF-induced mitogenesis or chemotaxis, we conclude that alpha PDGF receptor-associated PI-3 kinase activity is not required for either of these major PDGF signalling functions.
一种生物学功能未知的磷脂酰肌醇-3(PI-3)激酶活性与含酪氨酸激酶的蛋白质相关联,包括配体刺激后的一些生长因子受体。在β血小板衍生生长因子(βPDGF)受体中,激酶插入结构域内特定酪氨酸残基的磷酸化是其与该酶相互作用所必需的。我们发现,在αPDGF受体的激酶插入结构域内,用苯丙氨酸替代酪氨酸残基731或742并不损害PDGF诱导的受体或体内底物磷脂酶C-γ的酪氨酸磷酸化。此外,对配体刺激的磷脂酰肌醇周转不受影响。然而,这两种损伤均显著损害受体与PI-3激酶的结合。在表达任一突变受体的PDGF刺激细胞中,抗磷酸酪氨酸抗体可回收的PI-3激酶也显著降低。由于这两种突变均未消除PDGF诱导的有丝分裂或趋化性,我们得出结论,αPDGF受体相关的PI-3激酶活性对于这两种主要的PDGF信号功能均非必需。