{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T02:03:21Z","timestamp":1791597801594,"version":"4.3.4"},"reference-count":32,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","award":["184951"],"award-info":[{"award-number":["184951"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","award":["10003696"],"award-info":[{"award-number":["10003696"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Adenylyl cyclases (ACs) convert ATP into the second messenger cAMP, thus directly influencing cellular signaling in response to a wide variety of stimuli. Despite their physiological importance, structural studies of isoform\u2010specific AC regulation are compounded by difficulties in AC expression and purification. Here, we designed a chimeric construct AC95, combining human AC9 as a molecular scaffold and incorporating the catalytic\u2013allosteric core of human AC5. Cryo\u2010EM analysis of AC95 at 3.5\u2009\u00c5 resolution revealed a state of AC95 bound to both ATP\u03b1S and forskolin, demonstrating that the chimera partially reproduces allosteric regulation by forskolin while retaining the structural features of the AC9 scaffold. Although AC95 chimera retained the ability to bind to and be activated by forskolin, it lost the ability to be autoinhibited by the C2b domain of AC9. Moreover, AC95 is insensitive to inhibition by AC5 inhibitors SQ22,536 and NKY80, due to either targeting a site distinct from the catalytic\u2013allosteric core of AC5, altered conformational dynamics or long\u2010range allosteric effects imposed by chimeric scaffold, or a requirement for additional AC5 features absent in AC95. Our results establish an approach for investigating isoform\u2010specific regulation of mammalian ACs by small molecules, offering a potential path for structure\u2010based drug discovery targeting distinct AC isoforms.<\/jats:p>","DOI":"10.1002\/pro.70801","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T12:51:37Z","timestamp":1789649497000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Adenylyl cyclase 9 as a molecular scaffold to dissect the mechanisms of membrane adenylyl cyclases"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8846-5755","authenticated-orcid":false,"given":"Ilayda","family":"Kantarci","sequence":"first","affiliation":[{"name":"Laboratory of Biomolecular Research Paul Scherrer Institute  Villigen Switzerland"},{"name":"Institute of Molecular Biology and Biophysics ETH Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoriwa","family":"Haoriwa","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Research Paul Scherrer Institute  Villigen Switzerland"},{"name":"Institute of Molecular Biology and Biophysics ETH Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0962-9433","authenticated-orcid":false,"given":"Volodymyr M.","family":"Korkhov","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Research Paul Scherrer Institute  Villigen Switzerland"},{"name":"Institute of Molecular Biology and Biophysics ETH Zurich  Zurich 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