Characterization of a membrane-associated, receptor and G-protein responsive phosphoinositide-specific phospholipase C from avian erythrocytes

• Published in: FEBS letters 1993-08, Vol.328 (1), p.174-182
• Main Authors: Ruiz-Larrea, Fernanda, Berrie, Christopher P.
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Calcium - pharmacology; Chromatography, Affinity; Chromatography, Ion Exchange; Cytoskeleton; Cytosol - enzymology; Electrophoresis, Polyacrylamide Gel; Enzyme Stability - drug effects; Enzymes and enzyme inhibitors; Erythrocyte Membrane - enzymology; Erythrocyte Membrane - ultrastructure; Fundamental and applied biological sciences. Psychology; G-protein; GTP-Binding Proteins - metabolism; Guanosine 5'-O-(3-Thiotriphosphate) - metabolism; Hydrolases; Molecular Weight; Phosphatidylinositols - metabolism; Phosphoinositide; Phospholipase-C; Purinergic receptor; Receptors, Purinergic - metabolism; Turkey erythrocyte; Turkeys; Type C Phospholipases - isolation & purification; Type C Phospholipases - metabolism
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(93)80988-7

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 P 2y 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 GTPγS and P 2y-purinergic receptor activation.