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GNG12  -  guanine nucleotide binding protein (G...

Bos taurus

 
 
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Disease relevance of GNG12

  • Pertussis toxin delayed the onset of inhibition by GTP gamma S but did not prevent the inhibitory effect. alpha o-GTP gamma S or alpha o-GDP had little effect on PLC activity. alpha i-GTP gamma S and alpha i-GDP produced a 15% inhibition of PLC activity [1].
  • Biochemical characterization of the purified ram p25 revealed that the profile of [35S]GTP gamma S-binding activity was almost the same as that of recombinant ram p25 expressed in E. coli [2].
 

High impact information on GNG12

  • Activation of IK.ACh by recombinant G alpha-GTP gamma S was observed, but this was probably due to release of GTP gamma S from the protein [3].
  • Release of bound [35S]GTP gamma S from ARF3 required the presence of both GEP and unlabeled GTP or GTP gamma S; GDP was much less effective [4].
  • In the presence of 0.6-0.8 mM MgCl2 and 1 mM EDTA, binding of guanosine 5'-[gamma[35S]thio]triphosphate ([35S]GTP gamma S) by ARF1 and ARF3 was equally high without and with GEP [4].
  • The strongest effect was observed for gamma S, followed by h gamma Srec, gamma E, gamma A-F, gamma D, gamma B. Moreover, fluorescence properties of alpha-crystallins in the presence of bound beta(LOW)-and gamma-crystallins indicated that the formation of beta(LOW)/alpha- or gamma/alpha-crystallin complexes involved various binding sites [5].
  • In each case, binding appeared specific as judged by competition of unlabeled G beta gamma with radiolabeled G beta gamma and inhibition of binding by antigenic peptide or G alpha-GDP, but not G alpha-GTP gamma S (guanosine 5'-3-O-(thio)triphosphate) [6].
 

Biological context of GNG12

  • The carboxyl-terminal G alpha tGTP gamma S binding site of P gamma is different from but adjacent to its PDE inhibitory site [7].
  • The alpha T.GDP complex also is capable of inhibiting the alpha T.GTP gamma S-stimulated cyclic GMP hydrolysis by the native PDE [8].
  • The inactive (GDP-bound) form of the alpha-subunits appears to be the preferred substrate, with the phosphorylation being significantly reduced in alpha o and alpha i upon the binding of guanosine 5'-O-thiotriphosphate (GTP gamma S) and completely eliminated in the pure alpha-GTP gamma S complex of transducin [9].
  • Phospholipid-bound ARFGTP gamma S can also stably interact with and activate the catalytic subunit of cholera toxin, suggesting that ARFGTP provides a membrane anchor for cholera toxin and thereby facilitates its access to membrane-bound substrates [10].
  • Examination of the kinetics of dissociation of guanosine 5'-3-O-(thio)triphosphate (GTP gamma S) suggests that G226A Gs alpha is incapable of assuming the conformation necessary for high affinity binding of Mg2+ to the alpha subunit-GTP gamma S complex [11].
 

Anatomical context of GNG12

  • Removal of PDE's membrane-binding domain by limited trypsin digestion eliminated both the binding of PDE to vesicles and the ability of PDE to be activated by T alpha-GTP gamma S and membranes [12].
  • Using [35S]guanosine 5'-O-(3-thiotriphosphate) ([35S]GTP gamma S) as the binding ligand, purified myelin from bovine brain was found to contain approximately half the binding activity of whole white matter (138 +/- 9 vs. 271 +/- 18 pmol/mg of protein) [13].
  • Salicylate (3 mM) and piroxicam (50 microM) reduced [35S]GTP gamma S binding to purified plasma membranes [65 +/- 3.7 and 75 +/- 5.3% (P < 0.003) of control, respectively] [14].
  • [35S]GTP gamma S-binding activity is detected in the cytosol fraction but this activity is one-sixth to one-eighth of that of the crude membrane fraction [15].
  • [35S]ATP gamma S binding to intact bovine aortic endothelial cells was competed by ATP gamma S (Kd: 1.0 +/- 0.5 microM; mean +/- S.D.), but not by 2-MeSATP and UTP, indicating that these binding sites are neither the P2Y nor the P2U receptors [16].
 

Associations of GNG12 with chemical compounds

  • The 2.2 A crystal structure of activated rod transducin, Gt alpha.GTP gamma S, shows the bound GTP gamma S molecule occluded deep in a cleft between a domain structurally homologous to small GTPases and a helical domain unique to heterotrimeric G proteins [17].
  • Phosphatidylcholine vesicles also will enhance the alpha T.GTP gamma S-stimulated PDE activity (1.5-2-fold) relative to that measured in the absence of a lipid milieu [18].
  • [35S]GTP gamma S binding to 22K G was inhibited by pretreatment with N-ethylmaleimide [19].
  • This potentiation by AS/7 cannot be attributed simply to an increase in the apparent affinity of alpha T.GTP gamma S for the effector enzyme, nor to an increased affinity of the enzyme for the substrate cyclic GMP [18].
  • Reinsertion of these partially purified receptor preparations into phospholipid vesicles reconstituted guanine nucleotide-sensitive high affinity agonist binding, agonist-promoted GTPase and 35S-labeled guanosine 5'-O-(thiotriphosphate) [( 35S]GTP gamma S) binding activity in these preparations [20].
 

Physical interactions of GNG12

  • In these animals we studied several highly polymorphic genetic systems such as haemoglobin, albumin, the BoLA Complex (class I and II) and the gamma S crystallin gene [21].
 

Other interactions of GNG12

  • When the AS/7 is papain-treated to yield monovalent antibody molecules, complexation between these monovalent antibodies and alpha T still occurs (as reflected by the ability of these antibodies to block rhodopsin-alpha T coupling); however, the potentiation of the alpha T.GTP gamma S-stimulated PDE activity is lost [18].
  • Receptor-catalyzed activation of Gq/11 by GTP gamma S, measured as the ability to activate purified phospholipase C-beta 1, paralleled receptor-catalyzed [35S]GTP gamma S binding [22].
 

Analytical, diagnostic and therapeutic context of GNG12

  • Unmyristoylated ARFGTP gamma S cosediments with phospholipid vesicles and totally binds to phospholipid-cholate micelles, as seen by gel filtration chromatography [10].
  • The basic strategy was that the materials were initially incubated with [35S]GTP gamma S at 37 degrees C. After 4 degrees C incubation in the wells of an ELISA plate precoated with G-protein antibodies, the radioactivity of each well was counted [23].
  • Purified rhodopsin is recombined with a G-protein containing extract in the presence of [35S]GTP gamma S and after incubation the reaction is stopped by rapid cooling and filtration [24].

References

  1. G protein-mediated inhibition of phospholipase C activity in a solubilized membrane preparation. Litosch, I., Sulkholutskaya, I., Weng, C. J. Biol. Chem. (1993) [Pubmed]
  2. Identification and localization of gene expression of a low M(r) GTP-binding protein, ram p25 in pituitary gland. Nagata, K., Sakagami, H., Kondo, H., Nozawa, Y. Biochem. Biophys. Res. Commun. (1995) [Pubmed]
  3. Recombinant G-protein beta gamma-subunits activate the muscarinic-gated atrial potassium channel. Wickman, K.D., Iñiguez-Lluhl, J.A., Davenport, P.A., Taussig, R., Krapivinsky, G.B., Linder, M.E., Gilman, A.G., Clapham, D.E. Nature (1994) [Pubmed]
  4. Purification and characterization of a guanine nucleotide-exchange protein for ADP-ribosylation factor from spleen cytosol. Tsai, S.C., Adamik, R., Moss, J., Vaughan, M. Proc. Natl. Acad. Sci. U.S.A. (1996) [Pubmed]
  5. Subunit exchange demonstrates a differential chaperone activity of calf alpha-crystallin toward beta LOW- and individual gamma-crystallins. Putilina, T., Skouri-Panet, F., Prat, K., Lubsen, N.H., Tardieu, A. J. Biol. Chem. (2003) [Pubmed]
  6. G beta gamma binds directly to the G protein-gated K+ channel, IKACh. Krapivinsky, G., Krapivinsky, L., Wickman, K., Clapham, D.E. J. Biol. Chem. (1995) [Pubmed]
  7. The carboxyl terminus of the gamma-subunit of rod cGMP phosphodiesterase contains distinct sites of interaction with the enzyme catalytic subunits and the alpha-subunit of transducin. Skiba, N.P., Artemyev, N.O., Hamm, H.E. J. Biol. Chem. (1995) [Pubmed]
  8. The regulation of the cyclic GMP phosphodiesterase by the GDP-bound form of the alpha subunit of transducin. Kroll, S., Phillips, W.J., Cerione, R.A. J. Biol. Chem. (1989) [Pubmed]
  9. Insulin-dependent phosphorylation of GTP-binding proteins in phospholipid vesicles. Krupinski, J., Rajaram, R., Lakonishok, M., Benovic, J.L., Cerione, R.A. J. Biol. Chem. (1988) [Pubmed]
  10. Myristoylation is not required for GTP-dependent binding of ADP-ribosylation factor ARF1 to phospholipids. Franco, M., Chardin, P., Chabre, M., Paris, S. J. Biol. Chem. (1993) [Pubmed]
  11. The G226A mutant of Gs alpha highlights the requirement for dissociation of G protein subunits. Lee, E., Taussig, R., Gilman, A.G. J. Biol. Chem. (1992) [Pubmed]
  12. Membrane stimulation of cGMP phosphodiesterase activation by transducin: comparison of phospholipid bilayers to rod outer segment membranes. Malinski, J.A., Wensel, T.G. Biochemistry (1992) [Pubmed]
  13. Detection of G proteins in purified bovine brain myelin. Larocca, J.N., Golly, F., Ledeen, R.W. J. Neurochem. (1991) [Pubmed]
  14. Inhibition of neutrophil function by aspirin-like drugs (NSAIDS): requirement for assembly of heterotrimeric G proteins in bilayer phospholipid. Abramson, S.B., Leszczynska-Piziak, J., Clancy, R.M., Philips, M., Weissmann, G. Biochem. Pharmacol. (1994) [Pubmed]
  15. Multiple small molecular weight GTP-binding proteins in bovine brain cytosol. Purification and characterization of a 24KDa protein. Yamamoto, K., Kim, S., Kikuchi, A., Takai, Y. Biochem. Biophys. Res. Commun. (1988) [Pubmed]
  16. Evidence that most high-affinity ATP binding sites on aortic endothelial cells and membranes do not correspond to P2 receptors. Motte, S., Swillens, S., Boeynaems, J.M. Eur. J. Pharmacol. (1996) [Pubmed]
  17. The 2.2 A crystal structure of transducin-alpha complexed with GTP gamma S. Noel, J.P., Hamm, H.E., Sigler, P.B. Nature (1993) [Pubmed]
  18. An antibody-induced enhancement of the transducin-stimulated cyclic GMP phosphodiesterase activity. Phillips, W.J., Trukawinski, S., Cerione, R.A. J. Biol. Chem. (1989) [Pubmed]
  19. Small molecular weight GTP-binding proteins in human platelet membranes. Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000. Ohmori, T., Kikuchi, A., Yamamoto, K., Kim, S., Takai, Y. J. Biol. Chem. (1989) [Pubmed]
  20. The D2-dopamine receptor of anterior pituitary is functionally associated with a pertussis toxin-sensitive guanine nucleotide binding protein. Senogles, S.E., Benovic, J.L., Amlaiky, N., Unson, C., Milligan, G., Vinitsky, R., Spiegel, A.M., Caron, M.G. J. Biol. Chem. (1987) [Pubmed]
  21. An attempt to identify genetic markers of resistance or susceptibility to dermatophilosis in the zebu Brahman population of Martinique. Maillard, J.C., Palin, C., Trap, I., Bensaid, A. Revue d'élevage et de médecine vétérinaire des pays tropicaux. (1993) [Pubmed]
  22. Reconstitution of agonist-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis using purified m1 muscarinic receptor, Gq/11, and phospholipase C-beta 1. Berstein, G., Blank, J.L., Smrcka, A.V., Higashijima, T., Sternweis, P.C., Exton, J.H., Ross, E.M. J. Biol. Chem. (1992) [Pubmed]
  23. Measurement of GTP gamma S binding to specific G proteins in membranes using G-protein antibodies. Okamoto, T., Ikezu, T., Murayama, Y., Ogata, E., Nishimoto, I. FEBS Lett. (1992) [Pubmed]
  24. Investigation of rhodopsin catalyzed G-protein GTP-binding using [35S] GTP gamma S--effects of regeneration and hydroxylamine. Cook, N.J., Pellicone, C., Virmaux, N. Biochem. Int. (1985) [Pubmed]
 
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