Divergent regulation of phosphatidylinositol 3-kinase P85 alpha and P85 beta isoforms upon T cell activation.
Phosphatidylinositol (PtdIns) 3-kinase is composed of a catalytic p110 subunit and a regulatory p85 subunit. A synthetic phosphopeptide corresponding to the kinase insert of the human PDGF beta subunit receptor and monoclonal antibodies raised against the two described p85 isoforms, p85 alpha and p85 beta were used to isolate PtdIns 3-kinase from human T lymphocytes. We demonstrate that T cells express both p85 alpha and p85 beta proteins. Both isoforms tightly associate with a p110 protein and with PtdIns 3-kinase activity in T cells. Upon triggering of the T cell antigen receptor (TCR)/CD3 complex or activation of protein kinase C (PKC) the p110 protein complexed to p85 alpha becomes rapidly phosphorylated exclusively on serine residues. p85 alpha does not appear to undergo a change in its basal serine phosphorylation during T cell activation. In contrast, stimulation of the TCR/CD3 complex or PKC, results in a marked and rapid increase in phosphorylation of p85 beta on threonine residues. These data show that PtdIns 3-kinase can be a substrate for serine/threonine kinases in T cells. The differential phosphorylation of p85 alpha and p85 beta reveals the potential for divergent regulation and function of these two PtdIns 3-kinase isoforms during T cell activation.
Lymphocyte Activation Laboratory, Imperial Cancer Research Fund, London, United Kingdom.
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D000818:Animals; D017252:CD3 Complex / Q000378:metabolism; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008213:Lymphocyte Activation; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009036:Moths; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010710:Phosphates / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D010770:Phosphotransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013601:T-Lymphocytes / Q000201:enzymology* / Q000276:immunology; D014162:Transfection
D017252:CD3 Complex; D007527:Isoenzymes; D046911:Macromolecular Substances; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D013462:Sulfur Radioisotopes; D015240:Phorbol 12,13-Dibutyrate; D012694:Serine; D008715:Methionine; D010770:Phosphotransferases; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C