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PMID: 9789008 Published · ppublish English Journal Article

The helical domain of a G protein alpha subunit is a regulator of its effector.

Liu W, Northup JK

Abstract

The alpha subunit (Galpha) of heterotrimeric G proteins is a major determinant of signaling selectivity. The Galpha structure essentially comprises a GTPase "Ras-like" domain (RasD) and a unique alpha-helical domain (HD). We used the vertebrate phototransduction model to test for potential functions of HD and found that the HD of the retinal transducin Galpha (Galphat) and the closely related gustducin (Galphag), but not Galphai1, Galphas, or Galphaq synergistically enhance guanosine 5'-gamma[-thio]triphosphate bound Galphat (GalphatGTPgammaS) activation of bovine rod cGMP phosphodiesterase (PDE). In addition, both HDt and HDg, but not HDi1, HDs, or HDq attenuate the trypsin-activated PDE. GalphatGDP and HDt attenuation of trypsin-activated PDE saturate with similar affinities and to an identical 38% of initial activity. These data suggest that interaction of intact Galphat with the PDE catalytic core may be caused by the HD moiety, and they indicate an independent site(s) for the HD moiety of Galphat within the PDE catalytic core in addition to the sites for the inhibitory Pgamma subunits. The HD moiety of GalphatGDP is an attenuator of the activated catalytic core, whereas in the presence of activated GalphatGTPgammaS the independently expressed HDt is a potent synergist. Rhodopsin catalysis of Galphat activation enhances the PDE activation produced by subsaturating levels of Galphat, suggesting a HD-moiety synergism from a transient conformation of Galphat. These results establish HD-selective regulations of vertebrate retinal PDE, and they provide evidence demonstrating that the HD is a modulatory domain. We suggest that the HD works in concert with the RasD, enhancing the efficiency of G protein signaling.

MeSH Terms
3',5'-Cyclic-GMP Phosphodiesterases/metabolism Amino Acid Sequence Animals Cattle Cloning, Molecular Crystallography, X-Ray Enzyme Activation GTP Phosphohydrolases/chemistry GTP-Binding Proteins/chemistry,isolation & purification,metabolism Guanosine 5'-O-(3-Thiotriphosphate)/metabolism Kinetics Macromolecular Substances Mice Molecular Sequence Data Protein Structure, Secondary Rats Recombinant Proteins/chemistry,isolation & purification,metabolism Retinal Rod Photoreceptor Cells/enzymology Sequence Alignment Sequence Homology, Amino Acid
Chemicals
Macromolecular Substances Recombinant Proteins Guanosine 5'-O-(3-Thiotriphosphate) 3',5'-Cyclic-GMP Phosphodiesterases GTP Phosphohydrolases GTP-Binding Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Liu W
Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, 5 Research Court, Rockville, MD 20850, USA.
Northup J K
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1998-10-27
Pages
12878-83
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC23639
Subset
IM
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