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Characterization of a membrane‐associated, receptor and G‐protein responsive phosphoinositide‐specific phospholipase C from avian erythrocytes
Author(s) -
Ruiz-Larrea Fernanda,
Berrie Christopher P.
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80988-7
Subject(s) - purinergic receptor , phospholipase c , gtp' , membrane , receptor , g protein , biochemistry , polyacrylamide gel electrophoresis , enzyme , chemistry , phospholipase , biology , microbiology and biotechnology
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 [ 3 H]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.