Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Oct 1;319(Pt 1):173–178. doi: 10.1042/bj3190173

G-protein inhibition of phospholipase C-beta 1 in membranes: role of G-protein beta gamma subunits.

I Litosch 1
PMCID: PMC1217751  PMID: 8870665

Abstract

Rat liver plasma membranes reconstituted with bovine brain phospholipase C beta 1 (PLC- beta 1) exhibit a dual regulation of PLC- beta 1 activity by G-proteins. Guanosine 5'-[gamma-thio]triphosphate (GTP[S]; 0.1 nM) produced a 20-25% inhibition of PLC- beta 1 activity within 7 min of incubation. The addition of vasopressin resulted in near-basal levels of activity in the presence of 0.1 nM GTP[S]. Clonidine had little effect on the net inhibition due to GTP[S]. A similar antagonism between carbachol and GTP[S] occurred in cerebral cortical membranes containing endogenous PLC- beta 1 activity. alpha 0/i-GDP (a mixture of GDP-liganded G0 alpha and Gi alpha) attenuated the GTP[S]-dependent inhibition of PLC- beta 1 whereas alpha 0/i-GTP[S] had no effect, suggesting an involvement of G-protein beta gamma subunits in the inhibition of PLC- beta 1. Low concentrations of beta gamma subunits inhibited PLC- beta 1 activity. Inhibition was followed by reversal to basal activity and onset of stimulation as the beta gamma concentration was increased. Inhibition by beta gamma was dependent on the presence of membranes. These results indicate that G-protein beta gamma subunits constitute a mechanism by which G-protein mediate a rapid and transient inhibition of PLC- beta 1.

Full Text

The Full Text of this article is available as a PDF (358.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berstein G., Blank J. L., Jhon D. Y., Exton J. H., Rhee S. G., Ross E. M. Phospholipase C-beta 1 is a GTPase-activating protein for Gq/11, its physiologic regulator. Cell. 1992 Aug 7;70(3):411–418. doi: 10.1016/0092-8674(92)90165-9. [DOI] [PubMed] [Google Scholar]
  2. Berstein G., Blank J. L., Smrcka A. V., Higashijima T., Sternweis P. C., Exton J. H., Ross E. M. Reconstitution of agonist-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis using purified m1 muscarinic receptor, Gq/11, and phospholipase C-beta 1. J Biol Chem. 1992 Apr 25;267(12):8081–8088. [PubMed] [Google Scholar]
  3. Blank J. L., Brattain K. A., Exton J. H. Activation of cytosolic phosphoinositide phospholipase C by G-protein beta gamma subunits. J Biol Chem. 1992 Nov 15;267(32):23069–23075. [PubMed] [Google Scholar]
  4. Boyer J. L., Waldo G. L., Harden T. K. Beta gamma-subunit activation of G-protein-regulated phospholipase C. J Biol Chem. 1992 Dec 15;267(35):25451–25456. [PubMed] [Google Scholar]
  5. Cherifi Y., Pigeon C., Le Romancer M., Bado A., Reyl-Desmars F., Lewin M. J. Purification of a histamine H3 receptor negatively coupled to phosphoinositide turnover in the human gastric cell line HGT1. J Biol Chem. 1992 Dec 15;267(35):25315–25320. [PubMed] [Google Scholar]
  6. Clapham D. E., Neer E. J. New roles for G-protein beta gamma-dimers in transmembrane signalling. Nature. 1993 Sep 30;365(6445):403–406. doi: 10.1038/365403a0. [DOI] [PubMed] [Google Scholar]
  7. Delahunty T. M., Cronin M. J., Linden J. Regulation of GH3-cell function via adenosine A1 receptors. Inhibition of prolactin release, cyclic AMP production and inositol phosphate generation. Biochem J. 1988 Oct 1;255(1):69–77. doi: 10.1042/bj2550069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  9. Goldsmith P., Backlund P. S., Jr, Rossiter K., Carter A., Milligan G., Unson C. G., Spiegel A. Purification of heterotrimeric GTP-binding proteins from brain: identification of a novel form of Go. Biochemistry. 1988 Sep 6;27(18):7085–7090. doi: 10.1021/bi00418a062. [DOI] [PubMed] [Google Scholar]
  10. Gutowski S., Smrcka A., Nowak L., Wu D. G., Simon M., Sternweis P. C. Antibodies to the alpha q subfamily of guanine nucleotide-binding regulatory protein alpha subunits attenuate activation of phosphatidylinositol 4,5-bisphosphate hydrolysis by hormones. J Biol Chem. 1991 Oct 25;266(30):20519–20524. [PubMed] [Google Scholar]
  11. Hepler J. R., Kozasa T., Smrcka A. V., Simon M. I., Rhee S. G., Sternweis P. C., Gilman A. G. Purification from Sf9 cells and characterization of recombinant Gq alpha and G11 alpha. Activation of purified phospholipase C isozymes by G alpha subunits. J Biol Chem. 1993 Jul 5;268(19):14367–14375. [PubMed] [Google Scholar]
  12. Higashijima T., Ferguson K. M., Sternweis P. C., Smigel M. D., Gilman A. G. Effects of Mg2+ and the beta gamma-subunit complex on the interactions of guanine nucleotides with G proteins. J Biol Chem. 1987 Jan 15;262(2):762–766. [PubMed] [Google Scholar]
  13. Jhon D. Y., Lee H. H., Park D., Lee C. W., Lee K. H., Yoo O. J., Rhee S. G. Cloning, sequencing, purification, and Gq-dependent activation of phospholipase C-beta 3. J Biol Chem. 1993 Mar 25;268(9):6654–6661. [PubMed] [Google Scholar]
  14. Katada T., Oinuma M., Kusakabe K., Ui M. A new GTP-binding protein in brain tissues serving as the specific substrate of islet-activating protein, pertussis toxin. FEBS Lett. 1987 Mar 23;213(2):353–358. doi: 10.1016/0014-5793(87)81521-1. [DOI] [PubMed] [Google Scholar]
  15. Litosch I. Guanine nucleotides mediate stimulatory and inhibitory effects on cerebral-cortical membrane phospholipase C activity. Biochem J. 1989 Jul 1;261(1):245–251. doi: 10.1042/bj2610245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Litosch I., Sulkholutskaya I., Weng C. G protein-mediated inhibition of phospholipase C activity in a solubilized membrane preparation. J Biol Chem. 1993 Apr 25;268(12):8692–8697. [PubMed] [Google Scholar]
  17. Northup J. K., Sternweis P. C., Gilman A. G. The subunits of the stimulatory regulatory component of adenylate cyclase. Resolution, activity, and properties of the 35,000-dalton (beta) subunit. J Biol Chem. 1983 Sep 25;258(18):11361–11368. [PubMed] [Google Scholar]
  18. Park D., Jhon D. Y., Lee C. W., Lee K. H., Rhee S. G. Activation of phospholipase C isozymes by G protein beta gamma subunits. J Biol Chem. 1993 Mar 5;268(7):4573–4576. [PubMed] [Google Scholar]
  19. Pumiglia K. M., LeVine H., Haske T., Habib T., Jove R., Decker S. J. A direct interaction between G-protein beta gamma subunits and the Raf-1 protein kinase. J Biol Chem. 1995 Jun 16;270(24):14251–14254. doi: 10.1074/jbc.270.24.14251. [DOI] [PubMed] [Google Scholar]
  20. Ryu S. H., Cho K. S., Lee K. Y., Suh P. G., Rhee S. G. Purification and characterization of two immunologically distinct phosphoinositide-specific phospholipases C from bovine brain. J Biol Chem. 1987 Sep 15;262(26):12511–12518. [PubMed] [Google Scholar]
  21. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  22. Shaw K., Exton J. H. Identification in bovine liver plasma membranes of a Gq-activatable phosphoinositide phospholipase C. Biochemistry. 1992 Jul 14;31(27):6347–6354. doi: 10.1021/bi00142a026. [DOI] [PubMed] [Google Scholar]
  23. Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
  24. Smrcka A. V., Hepler J. R., Brown K. O., Sternweis P. C. Regulation of polyphosphoinositide-specific phospholipase C activity by purified Gq. Science. 1991 Feb 15;251(4995):804–807. doi: 10.1126/science.1846707. [DOI] [PubMed] [Google Scholar]
  25. Smrcka A. V., Sternweis P. C. Regulation of purified subtypes of phosphatidylinositol-specific phospholipase C beta by G protein alpha and beta gamma subunits. J Biol Chem. 1993 May 5;268(13):9667–9674. [PubMed] [Google Scholar]
  26. Sternweis P. C., Robishaw J. D. Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem. 1984 Nov 25;259(22):13806–13813. [PubMed] [Google Scholar]
  27. Taussig R., Tang W. J., Hepler J. R., Gilman A. G. Distinct patterns of bidirectional regulation of mammalian adenylyl cyclases. J Biol Chem. 1994 Feb 25;269(8):6093–6100. [PubMed] [Google Scholar]
  28. Taylor S. J., Chae H. Z., Rhee S. G., Exton J. H. Activation of the beta 1 isozyme of phospholipase C by alpha subunits of the Gq class of G proteins. Nature. 1991 Apr 11;350(6318):516–518. doi: 10.1038/350516a0. [DOI] [PubMed] [Google Scholar]
  29. Taylor S. J., Exton J. H. Guanine-nucleotide and hormone regulation of polyphosphoinositide phospholipase C activity of rat liver plasma membranes. Bivalent-cation and phospholipid requirements. Biochem J. 1987 Dec 15;248(3):791–799. doi: 10.1042/bj2480791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Taylor S. J., Smith J. A., Exton J. H. Purification from bovine liver membranes of a guanine nucleotide-dependent activator of phosphoinositide-specific phospholipase C. Immunologic identification as a novel G-protein alpha subunit. J Biol Chem. 1990 Oct 5;265(28):17150–17156. [PubMed] [Google Scholar]
  31. Ueda N., Iñiguez-Lluhi J. A., Lee E., Smrcka A. V., Robishaw J. D., Gilman A. G. G protein beta gamma subunits. Simplified purification and properties of novel isoforms. J Biol Chem. 1994 Feb 11;269(6):4388–4395. [PubMed] [Google Scholar]
  32. Uhing R. J., Prpic V., Jiang H., Exton J. H. Hormone-stimulated polyphosphoinositide breakdown in rat liver plasma membranes. Roles of guanine nucleotides and calcium. J Biol Chem. 1986 Feb 15;261(5):2140–2146. [PubMed] [Google Scholar]
  33. Wang Y., Durkin J. P. alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, but not N-methyl-D-aspartate, activates mitogen-activated protein kinase through G-protein beta gamma subunits in rat cortical neurons. J Biol Chem. 1995 Sep 29;270(39):22783–22787. doi: 10.1074/jbc.270.39.22783. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES