{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T15:19:43Z","timestamp":1781104783305,"version":"3.54.1"},"reference-count":42,"publisher":"National Academy of Sciences","issue":"2","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1999,1,19]]},"abstract":"<jats:p>\n                    Hormonal signals activate trimeric G proteins by substituting GTP for GDP bound to the G protein \u03b1 subunit (G\u03b1), thereby generating two potential signaling molecules, G\u03b1\u2013GTP and free G\u03b2\u03b3. The usefulness of dominant negative mutations for investigating Ras and other monomeric G proteins inspired us to create a functionally analogous dominant negative G\u03b1 mutation. Here we describe a mutant \u03b1 subunit designed to inhibit receptor-mediated hormonal activation of G\n                    <jats:sub>s<\/jats:sub>\n                    , the stimulatory regulator of adenylyl cyclase. To construct this mutant, we introduced into the \u03b1 subunit (\u03b1\n                    <jats:sub>s<\/jats:sub>\n                    ) of G\n                    <jats:sub>s<\/jats:sub>\n                    three separate mutations chosen because they impair \u03b1\n                    <jats:sub>s<\/jats:sub>\n                    function in complementary ways: the A\n                    <jats:sup>366<\/jats:sup>\n                    S mutant reduces affinity of \u03b1\n                    <jats:sub>s<\/jats:sub>\n                    for binding GDP, whereas the G\n                    <jats:sup>226<\/jats:sup>\n                    A and E\n                    <jats:sup>268<\/jats:sup>\n                    A mutations impair the protein\u2019s ability to bind GTP and to assume an active conformation. The triple mutant robustly inhibits (by up to 80%) G\n                    <jats:sub>s<\/jats:sub>\n                    -dependent hormonal stimulation of adenylyl cyclase in cultured cells. Inhibition is selective in that it does not affect cellular responses to expression of a constitutively active \u03b1\n                    <jats:sub>s<\/jats:sub>\n                    mutant (\u03b1\n                    <jats:sub>s<\/jats:sub>\n                    \u2013R\n                    <jats:sup>201<\/jats:sup>\n                    C) or to agonists for receptors that activate G\n                    <jats:sub>q<\/jats:sub>\n                    or G\n                    <jats:sub>i<\/jats:sub>\n                    . This \u03b1\n                    <jats:sub>s<\/jats:sub>\n                    triple mutant and cognate G\u03b1 mutants should provide specific tools for dissection of G protein-mediated signals in cultured cells and transgenic animals.\n                  <\/jats:p>","DOI":"10.1073\/pnas.96.2.499","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:39:15Z","timestamp":1027679955000},"page":"499-504","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":59,"title":["A G\n                    <sub>s\u03b1<\/sub>\n                    mutant designed to inhibit receptor signaling through G\n                    <sub>s<\/sub>"],"prefix":"10.1073","volume":"96","author":[{"given":"Taroh","family":"Iiri","sequence":"first","affiliation":[{"name":"Departments of Cellular and Molecular Pharmacology and Medicine and Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and Fourth Department of Internal Medicine, University of Tokyo School of Medicine, 3-28-6 Mejirodai, Bunkyo-ku, Tokyo 112, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sean M.","family":"Bell","sequence":"additional","affiliation":[{"name":"Departments of Cellular and Molecular Pharmacology and Medicine and Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and Fourth Department of Internal Medicine, University of Tokyo School of Medicine, 3-28-6 Mejirodai, Bunkyo-ku, Tokyo 112, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J.","family":"Baranski","sequence":"additional","affiliation":[{"name":"Departments of Cellular and Molecular Pharmacology and Medicine and Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and Fourth Department of Internal Medicine, University of Tokyo School of Medicine, 3-28-6 Mejirodai, Bunkyo-ku, Tokyo 112, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshiro","family":"Fujita","sequence":"additional","affiliation":[{"name":"Departments of Cellular and Molecular Pharmacology and Medicine and Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and Fourth Department of Internal Medicine, University of Tokyo School of Medicine, 3-28-6 Mejirodai, Bunkyo-ku, Tokyo 112, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry R.","family":"Bourne","sequence":"additional","affiliation":[{"name":"Departments of Cellular and Molecular Pharmacology and Medicine and Cardiovascular Research Institute, University of California, San Francisco, CA 94143; and Fourth Department of Internal Medicine, University of Tokyo School of Medicine, 3-28-6 Mejirodai, Bunkyo-ku, Tokyo 112, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1999,1,19]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.2.669"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/348125a0"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.56.070187.003151"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/27831"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.66.1.639"},{"key":"e_1_3_3_6_2","first-page":"134","volume-title":"Current Opinion in Cell Biology","author":"Bourne H R","year":"1997","unstructured":"H R Bourne Current Opinion in Cell Biology, eds L C Cantley, S R Coughlin (Current Biology, London) 9, No. 2, 134\u2013142 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