A monoclonal antibody against the rod outer segment guanyl nucleotide-binding protein, transducin, blocks the stimulatory and inhibitory G proteins of adenylate cyclase.

Hamm HE, Deretic D, Mazzoni MR, Moore CA, Takahashi JS, Rasenick MM
J Biol Chem. 1989 264 (19): 11475-82

PMID: 2544596

GTP-binding proteins have been implicated as transducers of a variety of biological signaling processes. These proteins share considerable structural as well as functional homology. Due to these similarities, it was thought that a monoclonal antibody that inhibits the light activation of the rod outer segment GTP-binding protein, tranducin (Gt), might exert some functional effect upon the G proteins that regulate the adenylate cyclase system. Antibody 4A, raised against the alpha subunit of Gt, cross-reacted (by hybridization on nitrocellulose) with purified alpha subunits of other G proteins (Gi and Gs, regulatory guanyl nucleotide-binding proteins that mediate inhibition and stimulation of adenylate cyclase, respectively) as long as they were not denatured. This antibody, which interferes with rod outer segment cGMP phosphodiesterase activation by blocking interaction between rhodopsin and Gt, also interfered with actions of both the stimulatory and inhibitory G proteins of adenylate cyclase from rat cerebral cortex membranes. Effects of monoclonal antibody (mAb) 4A were dose-dependent and not reversed by washing. mAb 4A also blocked the Gi-mediated inhibition of adenylate cyclase in the cyc- variant of S49 lymphoma and in doing so raised the level of adenylate cyclase activity in both the cyc- variant and the S49 wild type. There was no effect of mAb 4A on adenylate cyclase activity of the resolved catalytic subunit. These results suggest that the well known sequence homologies among the G proteins involved in cellular signal transduction may extend to the sites that interact with other members of signal-transducing cascades (receptors and effector molecules). Therefore, antibody 4A may serve as a useful tool to probe the similarities and differences among the various systems.

MeSH Terms (24)

3',5'-Cyclic-GMP Phosphodiesterases Adenylyl Cyclase Inhibitors Adenylyl Cyclases Animals Antibodies, Monoclonal Antibody Specificity Cell Membrane Cerebral Cortex Enzyme Activation Guanosine 5'-O-(3-Thiotriphosphate) Guanosine Triphosphate Guanylyl Imidodiphosphate Immunoblotting Light Lymphoma Mice Photoreceptor Cells Pineal Gland Rats Rhodopsin Rod Cell Outer Segment Thionucleotides Transducin Tumor Cells, Cultured

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