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

H. E. Hamm, D. Deretic, M. R. Mazzoni, C. A. Moore, J. S. Takahashi, M. M. Rasenick

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

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, transducin (G(t)), might exert some functional effect upon the G proteins that regulate the adenylate cyclase system. Antibody 4A, raised against the α subunit of G(t), cross-reacted (by hybridization on nitrocellulose) with purified α subunits of other G proteins (G(i) and G(s), 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 G(t), 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 G(i)-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.

Original languageEnglish (US)
Pages (from-to)11475-11482
Number of pages8
JournalJournal of Biological Chemistry
Volume264
Issue number19
StatePublished - Jul 28 1989

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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