Modification of AlF-4- and receptor-stimulated phospholipase C activity by G-protein beta gamma subunits.
Turkey erythrocyte membranes possess a phospholipase C that is markedly activated by P2Y-purinergic receptor agonists and guanine nucleotides. Reconstitution of [3H]inositol-labeled turkey erythrocyte membranes with guanine nucleotide regulatory protein (G-protein) beta gamma subunits resulted in inhibition of both AlF-4-stimulated adenylate cyclase and AlF-4-stimulated phospholipase C activities. The apparent potency (K0.5 approximately 1 microgram or 20 pmol of beta gamma/mg of membrane protein) of beta gamma subunits for inhibition of each enzyme activity was similar and occurred with beta gamma purified by different methodologies from turkey erythrocyte, bovine brain, or human placenta membranes. In contrast to the effect on AlF-4-stimulated activity, the stimulatory effect on phospholipase C of the P2Y-purinergic receptor agonist 2-methylthioadenosine 5'-triphosphate in the presence of guanine nucleotides was potentiated by 50-100% in a concentration-dependent manner by reconstitution of beta gamma subunits. beta gamma subunits did not affect the K0.5 value of 2-methylthioadenosine 5'-triphosphate for the stimulation of phospholipase C activity. These results indicate that beta gamma subunits influence phospholipase C activity in a concentration range similar to that necessary for regulation of adenylate cyclase activity and suggest the involvement of a G-protein possessing an alpha beta gamma heterotrimeric structure in coupling hormone receptors to phospholipase C.
Boyer
J L
JL
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill 27599.
Waldo
G L
GL
Evans
T
T
Northup
J K
JK
Downes
C P
CP
Harden
T K
TK
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000262:Adenylyl Cyclases / Q000097:blood; D000535:Aluminum / Q000494:pharmacology*; D017607:Aluminum Compounds; D000818:Animals; D001921:Brain / Q000378:metabolism; D002417:Cattle; D005576:Colforsin / Q000494:pharmacology; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000378:metabolism; D005260:Female; D005459:Fluorides; D005461:Fluorine / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011983:Receptors, Purinergic / Q000187:drug effects / Q000502:physiology*; D013873:Thionucleotides / Q000494:pharmacology*; D014422:Turkeys; D010738:Type C Phospholipases / Q000097:blood*
D017607:Aluminum Compounds; D046911:Macromolecular Substances; D011983:Receptors, Purinergic; D013873:Thionucleotides; D005576:Colforsin; C050992:tetrafluoroaluminate; D005461:Fluorine; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D000535:Aluminum; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D005459:Fluorides; C008501:2-methylthio-ATP