{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T06:28:09Z","timestamp":1779344889262,"version":"3.51.4"},"reference-count":46,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5340","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1997,11,7]]},"abstract":"<jats:p>\n            Palmitoylation of the \u03b1 subunit of the guanine nucleotide-binding protein G\n            <jats:sub>z<\/jats:sub>\n            inhibited by more than 90 percent its response to the guanosine triphosphatase (GTPase)\u2013accelerating activity of G\n            <jats:sub>z<\/jats:sub>\n            GAP, a G\n            <jats:sub>z<\/jats:sub>\n            -selective member of the regulators of G-protein signaling (RGS) protein family of GTPase-activating proteins (GAPs). Palmitoylation both decreased the affinity of G\n            <jats:sub>z<\/jats:sub>\n            GAP for the GTP-bound form of G\u03b1\n            <jats:sub>z<\/jats:sub>\n            by at least 90 percent and decreased the maximum rate of GTP hydrolysis. Inhibition was reversed by removal of the palmitoyl group by dithiothreitol. Palmitoylation of G\u03b1\n            <jats:sub>z<\/jats:sub>\n            also inhibited its response to the GAP activity of G\u03b1-interacting protein (GAIP), another RGS protein, and palmitoylation of G\u03b1\n            <jats:sub>i1<\/jats:sub>\n            inhibited its response to RGS4. The extent of inhibition of G\n            <jats:sub>z<\/jats:sub>\n            GAP, GAIP, RGS4, and RGS10 correlated roughly with their intrinsic GAP activities for the G\u03b1 target used in the assay. Reversible palmitoylation is thus a major determinant of G\n            <jats:sub>z<\/jats:sub>\n            deactivation after its stimulation by receptors, and may be a general mechanism for prolonging or potentiating G-protein signaling.\n          <\/jats:p>","DOI":"10.1126\/science.278.5340.1132","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:45:15Z","timestamp":1027763115000},"page":"1132-1135","source":"Crossref","is-referenced-by-count":135,"title":["Inhibition of Brain G\n            <sub>z<\/sub>\n            GAP and Other RGS Proteins by Palmitoylation of G Protein \u03b1 Subunits"],"prefix":"10.1126","volume":"278","author":[{"given":"Yaping","family":"Tu","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235\u20139041, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235\u20139041, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elliott M.","family":"Ross","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235\u20139041, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1074\/jbc.270.2.503","volume":"270","author":"Wedegaertner P. B.","year":"1995","unstructured":"Wedegaertner P. B., Wilson P. T., Bourne H. R., J. Biol. Chem. 270, 503 (1995);","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_2_3","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/S0955-0674(97)80056-7","volume":"9","author":"Mumby S. M.","year":"1997","unstructured":"Mumby S. M., Curr. Opin. Cell Biol. 9, 148 (1997);","journal-title":"Curr. Opin. Cell Biol."},{"key":"e_1_3_1_2_4","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/S0960-9822(95)00026-1","volume":"5","author":"Ross E. M.","year":"1995","unstructured":"Ross E. M., Curr. Biol. 5, 107 (1995);","journal-title":"Curr. Biol."},{"key":"e_1_3_1_2_5","doi-asserted-by":"crossref","unstructured":"; G. Milligan et al. Biochem. Soc. Trans. 23 583 (1995); C. Kleuss and A. G. Gilman Proc. Natl. Acad. Sci. U.S.A. 94 6116 (1997).","DOI":"10.1073\/pnas.94.12.6116"},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"3675","DOI":"10.1073\/pnas.90.8.3675","volume":"90","author":"Linder M. E.","year":"1993","unstructured":"Linder M. E., et al., Proc. Natl. Acad. Sci. U.S.A. 90, 3675 (1993).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_4_2","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1016\/0092-8674(94)90445-6","volume":"77","author":"Wedegaertner P. B.","year":"1994","unstructured":"Wedegaertner P. B., Bourne H. R., Cell 77, 1063 (1994).","journal-title":"Cell"},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"2800","DOI":"10.1073\/pnas.91.7.2800","volume":"91","author":"Mumby S. M.","year":"1994","unstructured":"Mumby S. M., Kleuss C., Gilman A. G., Proc. Natl. Acad. Sci. U.S.A. 91, 2800 (1994).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_6_2","first-page":"738","volume":"11","author":"Stanislaus D.","year":"1997","unstructured":"Stanislaus D., Jovanovick J. A., Brothers S., Conn P. M., Mol. Endocrinol. 11, 738 (1997).","journal-title":"Mol. Endocrinol."},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"23769","DOI":"10.1016\/S0021-9258(20)80448-7","volume":"268","author":"Degtyarev M. Y.","year":"1993","unstructured":"Degtyarev M. Y., Spiegel A. M., Jones T. L. Z., J. Biol. Chem. 268, 23769 (1993);","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_7_3","doi-asserted-by":"crossref","unstructured":"; ibid. 269 30898 (1994).","DOI":"10.1016\/S0021-9258(18)47366-8"},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"14592","DOI":"10.1073\/pnas.93.25.14592","volume":"93","author":"Iiri T.","year":"1996","unstructured":"Iiri T., Backlund P. S., Jones T. L. Z., Wedegaertner P. B., Bourne H. R., Proc. Natl. Acad. Sci. U.S.A. 93, 14592 (1996).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1042\/bj3020913","volume":"302","author":"Grassie M. A.","year":"1994","unstructured":"Grassie M. A., et al., Biochem. J. 302, 913 (1994);","journal-title":"Biochem. J."},{"key":"e_1_3_1_9_3","first-page":"717","volume":"313","author":"Galbiati F.","year":"1996","unstructured":"Galbiati F., Guzzi F., Magee A. I., Milligan G., Parenti M., ibid. 313, 717 (1996).","journal-title":"ibid."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"9667","DOI":"10.1074\/jbc.270.16.9667","volume":"270","author":"Wilson P. T.","year":"1995","unstructured":"Wilson P. T., Bourne H. R., J. Biol. Chem. 270, 9667 (1995).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_11_2","first-page":"496","volume":"271","author":"Hepler J. R.","year":"1996","unstructured":"Hepler J. R., et al., ibid. 271, 496 (1996).","journal-title":"ibid."},{"key":"e_1_3_1_12_2","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/0014-5793(94)01101-X","volume":"354","author":"Edgerton M. D.","year":"1994","unstructured":"Edgerton M. D., Chabert C., Chollet A., Arkinstall S., FEBS Lett. 354, 195 (1994);","journal-title":"FEBS Lett."},{"key":"e_1_3_1_12_3","doi-asserted-by":"crossref","first-page":"25001","DOI":"10.1016\/S0021-9258(19)74563-3","volume":"268","author":"Wedegaertner P. B.","year":"1993","unstructured":"Wedegaertner P. B., Chu D. H., Wilson P. T., Levis M. J., Bourne H. R., J. Biol. Chem. 268, 25001 (1993).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_13_2","doi-asserted-by":"crossref","first-page":"14806","DOI":"10.1021\/bi961846b","volume":"35","author":"Song J.","year":"1996","unstructured":"Song J., Dohlman H. G., Biochemistry 35, 14806 (1996).","journal-title":"Biochemistry"},{"key":"e_1_3_1_14_2","doi-asserted-by":"crossref","first-page":"3066","DOI":"10.1073\/pnas.85.9.3066","volume":"85","author":"Fong H. K. W.","year":"1988","unstructured":"Fong H. K. W., Yoshimoto K. K., Eversole-Cire P., Simon M. I., Proc. Natl. Acad. Sci. U.S.A. 85, 3066 (1988);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_14_3","first-page":"5384","volume":"85","author":"Matsuoka M.","year":"1988","unstructured":"Matsuoka M., Itoh H., Kozasa T., Kaziro Y., ibid. 85, 5384 (1988).","journal-title":"ibid."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","first-page":"2383","DOI":"10.1016\/S0021-9258(19)39988-0","volume":"265","author":"Casey P. J.","year":"1990","unstructured":"Casey P. J., Fong H. K. W., Simon M. I., Gilman A. G., J. Biol. Chem. 265, 2383 (1990).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_16_2","first-page":"5732","volume":"272","author":"Wang J.","year":"1997","unstructured":"Wang J., Tu Y., Woodson J., Song X., Ross E. M., ibid. 272, 5732 (1997).","journal-title":"ibid."},{"key":"e_1_3_1_17_2","unstructured":"Sequence data indicate that the purified brain G z GAP (15) is an RGS protein related to RET-RGS1 (31) and GAIP (26)."},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"3871","DOI":"10.1074\/jbc.272.7.3871","volume":"272","author":"Dohlman H. G.","year":"1997","unstructured":"Dohlman H. G., Thorner J., J. Biol. Chem. 272, 3871 (1997);","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_18_3","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/S0955-0674(97)80055-5","volume":"9","author":"Koelle M. R.","year":"1997","unstructured":"Koelle M. R., Curr. Opin. Cell Biol. 9, 143 (1997).","journal-title":"Curr. Opin. Cell Biol."},{"key":"e_1_3_1_19_2","doi-asserted-by":"crossref","first-page":"23594","DOI":"10.1074\/jbc.271.38.23594","volume":"271","author":"Duncan J. A.","year":"1996","unstructured":"Duncan J. A., Gilman A. G., J. Biol. Chem. 271, 23594 (1996).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_20_2","unstructured":"G\u03b1 z expressed in Sf9 cells is both myristoylated and palmitoylated (2) but palmitate is largely removed during purification. G\u03b1 z (4 \u03bcM) purified from Sf9 cells or myristoylated G \u03b1 i1 from E. coli (32) was palmitoylated by incubation for 2 to 3 hours at 30\u00b0C with 50 \u03bcM Pal-CoA either unlabeled or 3 H-labeled at 1000 cpm\/pmol in 50 mM NaHepes (pH 7.8) 2 mM EDTA and 7.5 mM CHAPS detergent (18). [ 3 H]Palmitoylation was measured by liquid scintillation counting after precipitation with 10% trichloroacetic acid (18) or SDS\u2013polyacrylamide gel electrophoresis (PAGE). The concentration of active G\u03b1 z was assayed according to the binding of 10 \u03bcM [ 35 S]GTP-\u03b3-S (15 33) except that the binding reaction mixture was incubated for 60 min in the presence of 25 mM (NH 4 ) 2 SO 4 which increases the rate of nucleotide exchange on G-protein \u03b1 subunits (34). Maximal observed binding of [ 35 S]GTP-\u03b3-S was 65 to 70% of that predicted from assay of total protein."},{"key":"e_1_3_1_21_2","unstructured":"Hydrolysis of G\u03b1 z -bound [\u03b3- 32 P]GTP and G z GAP activity were measured at 15\u00b0C as described (15). GAP activity is expressed either as the amount of bound GTP hydrolyzed above background at early times (quasi-linear time course) or as an apparent first-order rate constant ( k app ). A unit of GAP activity is defined as an increment in k app of 1 min \u20131 (15). Basal and stimulated hydrolysis of GTP bound to G\u03b1 i1 was measured with a single-turnover assay (15) that included centrifugal gel filtration to remove unbound [\u03b3- 32 P]GTP and [ 32 P]Pi formed during the binding reaction."},{"key":"e_1_3_1_22_2","unstructured":"To proteolyze G\u03b1 z near its NH 2 -terminus G\u03b1 z was first incubated with either 50 \u03bcM GTP-\u03b3-S (30\u00b0C for 90 min) or 50 \u03bcM guanosine diphosphate (GDP) 30 \u03bcM AlCl 3 10 mM NaF and 10 mM MgCl 2 (15\u00b0C for 20 min) in 25 mM NaHepes (pH 7.5) 1 mM EDTA 0.5 mM Mg 2+ 1 mM DTT and 0.1% Triton X-100. The mixture was further incubated with trypsin (0.05 milligrams per milligram of G\u03b1 z ) at 30\u00b0C for 30 min. Phenylmethylsulfonylfluoride (1 mM) and tosyl-lysyl-chloromethylketone (0.2 mM) were added and digestion was checked by SDS-PAGE. Trypsin cleaved activated G\u03b1 z after Arg 29 according to automated Edman sequencing of the large tryptic fragment (22)."},{"key":"e_1_3_1_23_2","unstructured":"Y. Tu and J. Wang unpublished data."},{"key":"e_1_3_1_24_2","unstructured":"Both reversal of inhibition by DTT treatment and mock palmitoylation of G\u03b1 z without Pal-CoA were routine controls."},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.1038\/383172a0"},{"key":"e_1_3_1_25_3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.2.428"},{"key":"e_1_3_1_26_2","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/S0092-8674(00)80117-8","volume":"86","author":"Berman D. M.","year":"1996","unstructured":"Berman D. M., Wilkie T. M., Gilman A. G., Cell 86, 445 (1996).","journal-title":"Cell"},{"key":"e_1_3_1_27_2","doi-asserted-by":"crossref","first-page":"11916","DOI":"10.1073\/pnas.92.25.11916","volume":"92","author":"De Vries L.","year":"1995","unstructured":"De Vries L., Mousli M., Wurmser A., Farquhar M. G., Proc. Natl. Acad. Sci. U.S.A. 92, 11916 (1995).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_28_2","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0092-8674(00)80998-8","volume":"84","author":"Koelle M. R.","year":"1996","unstructured":"Koelle M. R., Horvitz H. R., Cell 84, 115 (1996).","journal-title":"Cell"},{"key":"e_1_3_1_29_2","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/383175a0","volume":"383","author":"Hunt T. W.","year":"1996","unstructured":"Hunt T. W., Fields T. A., Casey P. J., Peralta E. G., Nature 383, 175 (1996).","journal-title":"Nature"},{"key":"e_1_3_1_30_2","unstructured":"Palmitoylation of G\u03b1 i1 also blocked acceleration by RGS4 of M2 muscarinic cholinergic receptor-stimulated steady-state GTPase activity when assayed in reconstituted phospholipid vesicles as described (15 22). The effect of palmitoylation does not therefore depend on interaction with detergent."},{"key":"e_1_3_1_31_2","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S0092-8674(00)80204-4","volume":"89","author":"Tesmer J. J. G.","year":"1997","unstructured":"Tesmer J. J. G., Berman D. M., Gilman A. G., Sprang S. R., Cell 89, 251 (1997);","journal-title":"Cell"},{"key":"e_1_3_1_31_3","unstructured":". Direct and specific contact of RGS proteins with Cys 3 of G\u03b1 is not crucial because mutation of this residue to Ala did not alter the responsiveness of G\u03b1 i1 to RGS4 (22)."},{"key":"e_1_3_1_32_2","doi-asserted-by":"crossref","first-page":"2945","DOI":"10.1073\/pnas.94.7.2945","volume":"94","author":"Faurobert E.","year":"1997","unstructured":"Faurobert E., Hurley J. B., Proc. Natl. Acad. Sci. U.S.A. 94, 2945 (1997).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_33_2","doi-asserted-by":"crossref","first-page":"4654","DOI":"10.1016\/S0021-9258(20)64372-1","volume":"266","author":"Linder M. E.","year":"1991","unstructured":"Linder M. E., et al., J. Biol. Chem. 266, 4654 (1991).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_34_2","first-page":"266","volume":"260","author":"Brandt D. R.","year":"1985","unstructured":"Brandt D. R., Ross E. M., ibid. 260, 266 (1985).","journal-title":"ibid."},{"key":"e_1_3_1_35_2","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/0076-6879(91)95165-G","volume":"195","author":"Ferguson K. M.","year":"1991","unstructured":"Ferguson K. M., Higashijima T., Methods Enzymol. 195, 188 (1991).","journal-title":"Methods Enzymol."},{"key":"e_1_3_1_36_2","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1074\/jbc.270.4.1734","volume":"270","author":"Kozasa T.","year":"1995","unstructured":"Kozasa T., Gilman A. G., J. Biol. Chem. 270, 1734 (1995).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_37_2","unstructured":"We thank A. Duncan for advice and [ 3 H]Pal-CoA D. Berman for RGS4 protein and for GAIP cDNA S. Popov for purified RGS10 P. Chidiac for advice K. Chapman and J. Woodson for expert technical assistance C. Slaughter and S. Afendis for peptide sequencing and M. Cobb S. Mukhopadhyay and S. Sprang for comments on the manuscript. Supported by NIH grant GM30355 R. A. Welch Foundation grant I-0982 and a postdoctoral fellowship to J.W. from Cadus Pharmaceuticals Corp."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.278.5340.1132","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T04:21:18Z","timestamp":1705119678000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.278.5340.1132"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,11,7]]},"references-count":46,"journal-issue":{"issue":"5340","published-print":{"date-parts":[[1997,11,7]]}},"alternative-id":["10.1126\/science.278.5340.1132"],"URL":"https:\/\/doi.org\/10.1126\/science.278.5340.1132","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,11,7]]}}}