Abstract
Regulator of G protein signaling (RGS) proteins are GTPase-activating proteins (GAP) for G protein α-subunits and are thought to be responsible for rapid deactivation of G protein mediated signaling pathway. In this present study, we demonstrate that PA is the most efficient candidate to inhibit GAP activity of RGS4. The functional significance of N-terminus of RGS4 in respose to PA-granted inhibition on GAP activity has been studied with the site mutation in the N-terminus of RGS4. These site-directed mutations in the N-terminal domain do not severely disrupt its association with liposomes of PA. However, RGS4L23E diminishes the inhibition of GAP activity by PA compared with the wild type RGS4, whereas RGSR22E abrogates the inhibitory effect by PA on GAP activity. The correspondent conformational discrepancy in the RGS domain of these mutants in the presence of PA vesicles was detected from fluorescence experiments. It is suggested that the functional pertinence between the N-terminus and RGS domain may be important to modulate PA-conferred inhibitory effect on its GAP activity.
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REFERENCES
Berman, D. M., Wilkie, R. M., and Gilman, A. G. (1996) Cell 86:445–452.
Bernstein, L. S., Grillo, A. A., Loranger, S. S., and Linder, M. E. (2000) J. Biol. Chem. 274:18520-18526.
Chen, C., Seow, K. T., Guo, K., Yaw, L. P., and Lin, S. (1999) J. Biol. Chem. 274:19799-19806.
Cunningham, M. L., Waldo, G. L., Hollinger, S., Hepler, J. R., and Harden, T. K. (2001) J. Biol. Chem. 276:5438-5444.
DeVries, L. and Farquhar, M. G. (1999) Trends Cell. Biol. 9:138-144.
Dunphy, J. T. and Linder, M. E. (1998) Biochim. Biophys. Acta 1436:245-261.
English, D., Cui, Y., and Siddiqui, R. A. (1996) Chem. Phys. Lipids 80:117-132.
Eftink, M. R. and Ghiron, C. A. (1981) Anal. Biochem. 114:199-227.
Ghosh, S., Strum, J. C., Sciorra, V. A., Daniel, L., and Bell, R. M. (1996) J. Biol. Chem. 271:8472-8480.
Hunt, T. W., Fields, T. A., Casey, P. J., and Peralta, E. G. (1996) Nature 383:172-175.
Kroon, A. I., Soekarjo, M. W., Gier, J. D., and Kruijff, B. D. (1990) Biochemistry 29:8229-8240.
Lakowicz, J. R. (1983) In: Principles of Fluorescence Spectroscopy, Plenum Press, New York.
Mumby, S. M. and Linder, M. E. (1994) Myristoylation of G-protein α-subunits. Methods Enzymol. 237:254-268.
Moy, F. J., Chanda, P. K., Cockett, M. I., Edris, W., Jones, P. G., Mason, K., Semus, S., and Powers, R. (2000) Biochemistry 39:7063-7073.
Natochin, M., Mcentaffer, R. L., and Artemyev, N. O. (1998) J. Biol. Chem. 273:6731-6735.
Neer, E. J. (1995) Cell 80:249-257.
Northup, J. K., Smigel, M. D., and Gilman, A. G. (1982) J. Biol. Chem. 257:11416-11423.
Ogier-Denis, E., Pattingre, S., Benna, J. E., and Codogno, P. (2000) J. Biol. Chem. 275:39090-39095.
Posner, B. A., Mukhopadhyay, S., Tesmer, J. J., Gilman, A. G., and Ross, E. M. (1999) Biochemistry 39:7773-7779.
Popov, S., Yu, K., Kozasa, T., and Wilkie, T. M. (1997) Proc. Natl. Acad. Sci. USA 94:7216-7220.
Ross, E. M. and Wilkie, T. M. (2000) Annu. Rev. Biochem. 69:795-827.
Srinivasa, S. P., Bernstein, L. S., Blumer, K. J., and Linder, M. E. (1998a) Proc. Natl. Acad. Sci. USA 95:5584-5589.
Srinivasa, S. P., Watson, N., Overton, M. C., and Blumer, K. J. (1998b) J. Biol. Chem. 273:1529-1533.
Tesmer, J. J., Berman, D. M., Gilman, A. G., and Sprang, S. R. (1997) Cell 89:251-261.
Tu, Y., Wang, J., and Ross, E. M. (1997) Science 278:1132-1135.
Tu, Y., Popov, S., Slaughter, C., and Ross, E. M. (1999) J. Biol. Chem. 274:38260-38267.
Wang, J., Ducret, A., Tu, Y., Kozaza, T., Aebesold, R., and Ross, E. M. (1998) J. Biol. Chem. 273:26014-26025.
Watson, N., Linder, M. E., Druey, K. M., Kehrl, J. H., and Blumer, K. J. (1996) Nature 383:172-175.
Zeng, W., Xu, X., Popov, S., Mukhopadhyay, S., Chidiac, P., Swistok, J., Danho, W., Yagaloff, K. A., Fisher, S. L., Ross, E. M., Muallem, S., and Wilkie, T. M. (1998) J. Biol. Chem. 273:34687-34690.
Zheng, B., Ma, Y. C., Ostrom, R. S., Lavoie, C., Gill, G. N., Insel, P. A., Huang, X. Y., and Farquhar, M. G. (2001) Science 294:1939-1942.
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Ou-Yang, YS., Tu, Y. & Yang, F. The Mutation in the N-Terminal Domain of RGS4 Disrupts PA-Conferred Inhibitory Effect on GAP Activity. Biosci Rep 23, 213–224 (2003). https://doi.org/10.1023/B:BIRE.0000007694.71158.02
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DOI: https://doi.org/10.1023/B:BIRE.0000007694.71158.02