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Volume 271, Number 43, Issue of October 25, 1996 pp. 26445-26448
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

COMMUNICATION:
Gbeta Subunit Interacts with a Peptide Encoding Region 956-982 of Adenylyl Cyclase 2
CROSS-LINKING OF THE PEPTIDE TO FREE Gbeta gamma BUT NOT THE HETEROTRIMER

(Received for publication, May 31, 1996, and in revised form, August 12, 1996)

Gezhi Weng Dagger , Jingrong Li Dagger , Jane Dingus , John D. Hildebrandt , Harel Weinstein Dagger par and Ravi Iyengar Dagger

From the Departments of Dagger  Pharmacology and par  Physiology and Biophysics, Mount Sinai School of Medicine, City University of New York, New York 10029 and the  Department of Pharmacology, Medical University of South Carolina, Charleston, South Carolina 29425

The region encoded by amino acids 956-982 of adenylyl cyclase 2 is important for Gbeta gamma stimulation. Interactions of a peptide encoding the 956-982 region of adenylyl cyclase 2 (QEHAQEPERQYMHIGTMVEFAYALVGK (QEHA peptide)) with Gbeta gamma subunits were studied. QEHA peptide was covalently attached to beta  subunit of free Gbeta gamma by the cross-linker N-succinimidyl(4-iodoacetyl)aminobenzoate. Cross-linking was proportional to the amount of QEHA peptide added; other control peptides cross-linked minimally. When Go was used, very little cross-linking was observed with GDP and EDTA, but upon activation by guanosine 5'-3-O-(thio)triphosphate and Mg2+, specific cross-linking of the QEHA peptide to Gbeta was observed. We conclude that beta  subunits of G proteins contain effector interaction domains that are occluded by Galpha subunits in the heterotrimer. Molecular modeling studies used to dock the QEHA peptide on to Gbeta indicate that amino acids 75-165 of Gbeta may be involved in effector interactions.


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