Phospholipase C-gamma 1 and phospholipase C-gamma 2 are substrates of the B cell antigen receptor associated protein tyrosine kinase.
Cross-linking the antigen receptor on B cells results in a rapid increase in protein tyrosine kinase activity as detected by increased phosphorylation on tyrosine residues of multiple proteins. Although the identity of most of this substrates remains unknown, some have been proposed. One possible substrate of the antigen receptor-associated kinase is phospholipase C (PLC). Since multiple isoforms of PLC have been identified, we have studied which isoforms are targets of the antigen receptor. PLC-gamma 1 and PLC-gamma 2 but not PLC-beta 1 or PLC-delta 1 were detected in human B cells. Immunoprecipitation with antibodies against PLC-gamma 1 or PLC-gamma 2 and subsequent Western blotting with anti-phosphotyrosine antibodies revealed that both PLC-gamma 1 and PLC-gamma 2 are tyrosine phosphorylated in stimulated but not in resting B cells. This was confirmed by experiments whereby B cell lysates were immunoprecipitated with anti-phosphotyrosine antibody and subsequently blotted with antibodies against PLC-gamma 1 or PLC-gamma 2. Further, the specific protein tyrosine kinase inhibitors, tyrphostins, which block phospholipase-C activation and proliferation of B cells also inhibited tyrosine phosphorylation on both PLC-gamma 1 and PLC-gamma 2. We conclude that both isoforms PLC-gamma 1 and PLC-gamma 2 are targets of the antigen receptor-associated protein tyrosine kinase.
Roifman
C M
CM
Division of Immunology/Allergy, Hospital for Sick Children, Toronto, Ontario, Canada.
Wang
G
G
D000888:Antibodies, Anti-Idiotypic; D001402:B-Lymphocytes / Q000378:metabolism*; D003429:Cross Reactions; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology; D007527:Isoenzymes / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D010738:Type C Phospholipases / Q000276:immunology / Q000378:metabolism*
D000888:Antibodies, Anti-Idiotypic; D007075:Immunoglobulin M; D007527:Isoenzymes; D011947:Receptors, Antigen, B-Cell; D011505:Protein-Tyrosine Kinases; D010738:Type C Phospholipases