{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:55:20Z","timestamp":1782280520185,"version":"3.54.5"},"posted":{"date-parts":[[2025,9,11]]},"group-title":"Biophysics","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T00:00:00Z","timestamp":1757548800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,9,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The \u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  adrenergic receptor (\u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  AR) is a clinically important target for various diseases including hypertension, diabetes and chronic pain. Here, using single-molecule fluorescence resonance energy transfer imaging, we show how agonist-specific activation dynamics in both structured transmembrane domain (TMD) and intrinsically disorders regions (IDRs) of \u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  AR lead to diverse signaling profiles. Through seven pairs of strategically designed fluorophore labels, we systematically investigate the real-time conformational changes of \u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  AR. Our study reveals unique TM6 dynamics in \u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  AR, featured by a high energy barrier for agonist-induced outward movements essential for activation. In contrast, we identify agonist-specific conformational dynamics of a partially disordered extracellular loop (ECL2), highlighting its role as a dynamic regulatory module that controls receptor function. Moreover, we characterize the conformational landscapes of the long third intracellular loop (ICL3), revealing its compact structural features and membrane-proximal localization in the basal state, where it acts as a negative allosteric regulator in transducer coupling. Furthermore, we identify multiful functional sub-states of ICL3 that are dynamically modulated by both kinase phosphorylation and drug efficacy. These findings offer previously underappreciated structural and dynamic insights into \u03b1\n                  <jats:sub>2A<\/jats:sub>\n                  AR function governed by both TMD and IDRs, and may open up new avenues for the development of better therapeutics.\n                <\/jats:p>","DOI":"10.1101\/2025.09.10.675278","type":"posted-content","created":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T21:05:47Z","timestamp":1757624747000},"source":"Crossref","is-referenced-by-count":1,"title":["The role of intrinsically disordered domains in regulating G protein coupled receptor signaling"],"prefix":"10.64898","author":[{"given":"Jun","family":"Xu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruoyi","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander M.","family":"Garces","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harald","family":"H\u00fcbner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dorothee","family":"Weikert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Gmeiner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael T.","family":"Lerch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian K.","family":"Kobilka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025091402500826000_2025.09.10.675278v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd.2017.178"},{"key":"2025091402500826000_2025.09.10.675278v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2023.102574"},{"key":"2025091402500826000_2025.09.10.675278v1.3","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2023.1121962"},{"key":"2025091402500826000_2025.09.10.675278v1.4","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15841"},{"key":"2025091402500826000_2025.09.10.675278v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2014.08.002"},{"key":"2025091402500826000_2025.09.10.675278v1.6","doi-asserted-by":"publisher","DOI":"10.3390\/biom13101431"},{"key":"2025091402500826000_2025.09.10.675278v1.7","doi-asserted-by":"publisher","DOI":"10.2344\/0003-3006-62.1.31"},{"key":"2025091402500826000_2025.09.10.675278v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejphar.2017.09.018"},{"key":"2025091402500826000_2025.09.10.675278v1.9","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1097\/00001503-200110000-00009","article-title":"Alpha2 receptors and agonists in pain management","volume":"14","year":"2001","journal-title":"Current Opinion in Anaesthesiology"},{"key":"2025091402500826000_2025.09.10.675278v1.10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.122368499"},{"key":"2025091402500826000_2025.09.10.675278v1.11","doi-asserted-by":"publisher","DOI":"10.1126\/science.abn7065"},{"key":"2025091402500826000_2025.09.10.675278v1.12","doi-asserted-by":"crossref","first-page":"4740","DOI":"10.1016\/S0021-9258(18)42894-3","article-title":"Sites in the third intracellular loop of the alpha 2A-adrenergic receptor confer short term agonist-promoted desensitization. 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