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Endocrinology Vol. 139, No. 11 4547-4555
Copyright © 1998 by The Endocrine Society

ARTICLES

Luteinizing Hormone/Choriogonadotropin Receptor-Mediated Activation of Heterotrimeric Guanine Nucleotide Binding Proteins in Ovarian Follicular Membranes1

Rajsree M. Rajagopalan-Gupta2, Marilyn L. G. Lamm3, Sutapa Mukherjee, Mark M. Rasenick and Mary Hunzicker-Dunn

Department of Cell and Molecular Biology (R.M.R.-G., M.L.G.L., S.M., M.H.-D.), Northwestern University Medical School, Chicago Illinois 60611; and Departments of Physiology and Biophysics and Psychiatry (M.M.R.), University of Illinois College of Medicine, Chicago, Illinois 60608

Address all correspondence and requests for reprints to: Dr. Mary Hunzicker-Dunn, Department of Cell and Molecular Biology, Northwestern University Medical School, 303 East Chicago Avenue, Chicago, Illinois 60611. E-mail: mhd{at}nwu.edu

The LH/CG receptor signals to adenylyl cyclase via the stimulatory heterotrimeric GTP binding regulatory protein, Gs, and to phospholipase C and potentially to other effectors, such as ion channels, via a G protein or proteins that have not been identified in gonadal cells. To identify G proteins activated in a physiological membrane environment upon LH/CG receptor activation, we used the ability of activated G proteins to bind GTP and incubated ovarian follicular membranes with the photoaffinity GTP analog, P3-(4-azidoanilido)-P1-5'-GTP ([32P]AAGTP). Results showed that human CG (hCG) stimulated the binding of [32P]AAGTP to a 45-kDa protein(s) in follicular membranes that comigrated with immunoreactive G{alpha}s, G{alpha}q/11, and G{alpha}13. When G{alpha} proteins were immunoprecipitated from Triton X-100 solubilized membrane extracts after photoaffinity labeling with [32P]AAGTP, a time-dependent increase in hCG-dependent [32P]AAGTP binding to G{alpha}s, G{alpha}q/11, and G{alpha}i was detected. hCG-dependent [32P]AAGTP binding to G{alpha}13 was also detected. These results demonstrate that agonist-dependent LH/CG receptor activation promotes the activation of Gs, Gi, Gq/11, and G13 in porcine ovarian follicular membranes. These results further suggest that G{alpha}s remains coupled to the agonist-bound LH/CG receptor during at least the initial 10 min after agonist-dependent receptor activation.




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