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PANX1 channel activity is regulated by phosphorylation at Y198, S205 and Y308 residues, but the physiological significance of these modifications is unknown. Here, we utilize newly developed PANX1 Y198F, S205A and Y308F phospho-dead mutant mice to test physiological changes related to hemodynamics. Radiotelemetry-measured blood pressure was decreased in Y198F, increased in Y308F, but unchanged in S205A mice at baseline. Clonidine-sensitive sympathetic-driven hypertension was observed in all mouse lines except Y198F. Pressure myography of third-order mesenteric arteries revealed \u03b1-adrenergic contractile responses were decreased in Y198F, slightly enhanced in Y308F, but unchanged in S205A, with responses in Y198F vessels mimicking controls treated with PANX1 inhibitors. To understand signaling changes driving these phenotypes, we performed mesenteric artery bulk RNA sequencing, but found a minimal number of differentially expressed genes between phospho-dead mutants and controls. Similarly, co-immunoprecipitation-mass spectrometry of wildtype or phospho-dead mutant-expressing vascular SMCs revealed few interacting proteins distinct to each PANX1 variant. However, PANX1 channel activity assessments in HEK293T cells expressing the \u03b11D-adrenergic receptor as well as each phospho-dead mutant PANX1 showed that phenylephrine-induced ATP release from Y198F channels was significantly decreased compared to wildtype, but current was unaffected. Conversely, basal and phenylephrine-induced S205A and Y308F currents were reduced, but ATP release resembled controls. Taken together, these findings indicate that distinct PANX1 phosphorylation determines PANX1 metabolite release versus current conducting properties and in turn, regulates physiological outcomes in the vasculature.<\/jats:p>\n                <jats:sec>\n                  <jats:title>One Sentence Summary<\/jats:title>\n                  <jats:p>PANX1 Y198 phosphorylation-mediated ATP release is a major driver of \u03b1-adrenergic vasoconstriction in vascular smooth muscle cells.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.12.738047","type":"posted-content","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T21:40:22Z","timestamp":1784324422000},"source":"Crossref","is-referenced-by-count":1,"title":["Pannexin 1 phosphorylation sites differentially modulate channel activity and physiological outcomes"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6025-8300","authenticated-orcid":false,"given":"Brooke L.","family":"O\u2019Donnell","sequence":"first","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke S.","family":"Dunaway","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"},{"name":"Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuexin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Skylar A.","family":"Loeb","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"},{"name":"Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzanna J.","family":"Ju\u015bkiewicz","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"},{"name":"Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wyatt J.","family":"Schug","sequence":"additional","affiliation":[{"name":"Robert M. 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Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"},{"name":"Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa A.","family":"Luse","sequence":"additional","affiliation":[{"name":"Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, Virginia, USA"},{"name":"Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha C.","family":"Bielefeld","sequence":"additional","affiliation":[{"name":"Robert M. 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Penuela , Pannexin 1 binds beta-catenin to modulate melanoma cell growth and metabolism. J Biol Chem, 100478 (2021).","DOI":"10.1016\/j.jbc.2021.100478"},{"key":"2026072106251353000_2026.07.12.738047v1.13","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1111\/febs.17396","article-title":"Pannexin 1 crosstalk with the Hippo pathway in malignant melanoma","volume":"292","year":"2025","journal-title":"FEBS J"},{"key":"2026072106251353000_2026.07.12.738047v1.14","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvs187"},{"key":"2026072106251353000_2026.07.12.738047v1.15","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.118.311290"},{"key":"2026072106251353000_2026.07.12.738047v1.16","doi-asserted-by":"crossref","unstructured":"M. Billaud , Y. Chiu , A. W. Lohman , T. Parpaite , J. T. Butcher , S. M. Mutchler , L. J. DeLalio , M. V. Artamonov , J. K. Sandilos , A. K. Best , A. V. Somlyo , R. J. Thompson , T. H. Le , K. S. Ravichandran , D. A. Bayliss , B. E. Isakson , A molecular signature in the pannexin1 intracellular loop confers channel activation by the a1 adrenoreceptor in smooth muscle cells. Sci Signal 8, (2015).","DOI":"10.1126\/scisignal.2005824"},{"key":"2026072106251353000_2026.07.12.738047v1.17","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.006982"},{"key":"2026072106251353000_2026.07.12.738047v1.18","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.110.237594"},{"key":"2026072106251353000_2026.07.12.738047v1.19","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1161\/HYPERTENSIONAHA.122.20280","article-title":"Amount of Pannexin 1 in Smooth Muscle Cells Regulates Sympathetic Nerve-Induced Vasoconstriction","volume":"80","year":"2023","journal-title":"Hypertension"},{"key":"2026072106251353000_2026.07.12.738047v1.20","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.312380"},{"key":"2026072106251353000_2026.07.12.738047v1.21","doi-asserted-by":"crossref","unstructured":"M. E. Good , S. A. 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