Characterization of a membrane-associated, receptor and G-protein responsive phosphoinositide-specific phospholipase C from avian erythrocytes.
We describe the reconstitution and purification of a membrane-associated phosphoinositide-specific phospholipase C (PIC) from turkey erythrocyte ghosts. This PIC is responsive to a G-protein coupled to P2y purinergic receptors which are expressed in turkey erythrocytes. Reconstitution is achieved by adding partially purified PIC to [3H]inositol-prelabelled turkey erythrocyte membranes depleted of their endogenous PIC (acceptor membranes). PIC activity is associated with a 52 kDa polypeptide on SDS-polyacrylamide gel electrophoresis. Addition of a 307-fold purified enzyme to the acceptor membranes has no effect on basal PIC activity, but markedly increases the response to GTP gamma S and P2y-purinergic receptor activation.
National Centre of Biotechnology, Cantoblanco, Madrid, Spain.
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Ruiz-Larrea
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Berrie
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D004910:Erythrocyte Membrane / Q000201:enzymology* / Q000648:ultrastructure; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D008970:Molecular Weight; D010716:Phosphatidylinositols / Q000378:metabolism; D011983:Receptors, Purinergic / Q000378:metabolism*; D014422:Turkeys; D010738:Type C Phospholipases / Q000302:isolation & purification / Q000378:metabolism*
D010716:Phosphatidylinositols; D011983:Receptors, Purinergic; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D010738:Type C Phospholipases; D019204:GTP-Binding Proteins; D002118:Calcium