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Identification of Critical Regions on Phospholipase C-βl Required for Activation by G-proteins

Wu, Dianqing and Jiang, Huiping and Katz, Arieh and Simon, Melvin I. (1993) Identification of Critical Regions on Phospholipase C-βl Required for Activation by G-proteins. Journal of Biological Chemistry, 268 (5). pp. 3704-3709. ISSN 0021-9258.

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In order to determine which portion of phosphoinositide-specific phospholipase C (PLC)-β1 is required for activation by Gαq, a series of specific deletions and truncations of PLC-β1 cDNA were prepared. After transfection of COS-7 cells with these cDNA clones, the activity and localization of the expressed proteins were determined. Specific deletions in the C-terminal end of the protein did not lead to loss of intrinsic enzymatic activity but did result in loss of the ability to be activated by Gαq. The region required for activation was localized to the amino acid sequence corresponding to residues 903-1142 of PLC-β1. This region was further subdivided into two sequences; one extending from residues Thr-903 to Gln-1030 that was required for particulate fraction association as well as for activation by Gαq and the other extending from residues Gln-1030 to Leu-1142 that was required for interaction with G α subunits. These results were confirmed by the observation that the C-terminal portion of PLC-β1, when co-expressed with the muscarinic acetylcholine receptor type 1 or the α1C-adrenergic receptor in COS-7 cells, markedly inhibited ligand-induced release of inositol phosphates. In an in vitro system, two peptides derived from the G-protein interaction region at the C terminus were found to inhibit the guanosine 5'-3-O-(thio)triphosphate-dependent activation of PLC-β1 by Gαq. This further localized the sites on PLC-β1 which are involved in interaction with G-protein α subunits.

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Additional Information:© 1993 American Society for Biochemistry and Molecular Biology, Inc. Received for publication, August 6, 1992. This work was supported by United States Public Health services Grant GM34236 (to M. I. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. We thank Dr. S. G. Rhee for the PLC-βI cDNA, PLC-βl antibodies, and purified PLC-β1 protein, Drs. John Knopf, Ron Kriz, and Lin-ling Lin for the PLC-β3 cDNA, and Dr. Chang-Ho Lee for assistance with PLC assay.
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United States Public Health Service (USPHS)GM34236
Record Number:CaltechAUTHORS:20120308-103110336
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Official Citation:Identification of critical regions on phospholipase C-beta 1 required for activation by G-proteins. D Wu, H Jiang, A Katz, and M I Simon J. Biol. Chem. 1993 268: 3704-3709.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29645
Deposited By: Ruth Sustaita
Deposited On:12 Mar 2012 20:36
Last Modified:23 Aug 2016 10:10

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