{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T19:16:14Z","timestamp":1789067774167,"version":"build-2803163510"},"reference-count":47,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T00:00:00Z","timestamp":1662422400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100004411","name":"Heart and Stroke Foundation of Canada","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100004411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Pharmacology Res &amp; Perspec"],"published-print":{"date-parts":[[2022,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:label\/><jats:p>Aldosterone exerts some of its effects not by binding to mineralocorticoid receptors, but rather by acting via G protein\u2010coupled estrogen receptors (GPER). To determine if aldosterone binds directly to GPER, we studied the ability of aldosterone to compete for the binding of [<jats:sup>3<\/jats:sup>H] 2\u2010methoxyestradiol ([<jats:sup>3<\/jats:sup>H] 2\u2010ME), a high potency GPER\u2010selective agonist. We used GPER gene transfer to engineer Sf9\u2010cultured insect cells to express GPER. We chose insect cells to avoid interactions with any intrinsic mammalian receptors for aldosterone. [<jats:sup>3<\/jats:sup>H] 2\u2010ME binding was saturable and reversible to a high\u2010affinity population of receptors with K<jats:sub>d<\/jats:sub>\u00a0=\u20093.7\u00a0nM and B<jats:sub>max<\/jats:sub>\u00a0=\u20092.2\u00a0pmol\/mg. Consistent with agonist binding to G Protein\u2010coupled receptors, [<jats:sup>3<\/jats:sup>H] 2\u2010ME high\u2010affinity state binding was reduced in the presence of the hydrolysis\u2010resistant GTP analog, GppNHp. [<jats:sup>3<\/jats:sup>H] 2\u2010ME binding was competed for by the GPER agonist G1, the GPER antagonist G15, estradiol (E2), as well as aldosterone (Aldo). The order of potency for competing for [<jats:sup>3<\/jats:sup>H] 2\u2010ME binding, namely 2ME\u2009&gt;\u2009Aldo\u2009&gt;\u2009E2\u2009\u2265\u2009G1, paralleled the orders of potency for inhibition of cell proliferation and inhibition of ERK phosphorylation by ligands acting at GPER. These data confirm the ability of aldosterone to interact with the GPER, consistent with the interpretation that aldosterone likely mediates its GPER\u2010dependent effects by direct binding to the GPER.<\/jats:p><\/jats:sec><jats:sec><jats:title>Significance <jats:bold>statement<\/jats:bold><\/jats:title><jats:p>Despite the growing evidence for aldosterone's actions via G protein\u2010coupled estrogen receptors (GPER), there remains significant skepticism that aldosterone can directly interact with GPER. The current studies are the first to demonstrate directly that aldosterone indeed is capable of binding to the GPER and thus likely mediates its GPER\u2010dependent effects by direct binding to the receptor.<\/jats:p><\/jats:sec>","DOI":"10.1002\/prp2.995","type":"journal-article","created":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T09:16:08Z","timestamp":1662455768000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Correlation of functional and radioligand binding characteristics of <scp>GPER<\/scp> ligands confirming aldosterone as a <scp>GPER<\/scp> agonist"],"prefix":"10.1002","volume":"10","author":[{"given":"Qingming","family":"Ding","sequence":"first","affiliation":[{"name":"Institute of Cardiovascular Sciences Albrechtsen Research Centre  Winnipeg 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