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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2022,8,2]]},"abstract":"<jats:p>In response to acidic pH, the widely expressed proton-activated chloride (PAC) channel opens and conducts anions across cellular membranes. By doing so, PAC plays an important role in both cellular physiology (endosome acidification) and diseases associated with tissue acidosis (acid-induced cell death). Despite the available structural information, how proton binding in the extracellular domain (ECD) leads to PAC channel opening remains largely unknown. Here, through comprehensive mutagenesis and electrophysiological studies, we identified several critical titratable residues, including two histidine residues (H130 and H131) and an aspartic acid residue (D269) at the distal end of the ECD, together with the previously characterized H98 at the transmembrane domain\u2013ECD interface, as potential pH sensors for human PAC. Mutations of these residues resulted in significant changes in pH sensitivity. Some combined mutants also exhibited large basal PAC channel activities at neutral pH. By combining molecular dynamics simulations with structural and functional analysis, we further found that the \u03b212 strand at the intersubunit interface and the associated \u201cjoint region\u201d connecting the upper and lower ECDs allosterically regulate the proton-dependent PAC activation. Our studies suggest a distinct pH-sensing and gating mechanism of this new family of ion channels sensitive to acidic environment.<\/jats:p>","DOI":"10.1073\/pnas.2200727119","type":"journal-article","created":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T15:21:05Z","timestamp":1658762465000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":34,"title":["Molecular determinants of pH sensing in the proton-activated chloride channel"],"prefix":"10.1073","volume":"119","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8676-4444","authenticated-orcid":false,"given":"James","family":"Osei-Owusu","sequence":"first","affiliation":[{"name":"Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ekaterina","family":"Kots","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10065"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4412-4916","authenticated-orcid":false,"given":"Zheng","family":"Ruan","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, Van Andel Institute, Grand Rapids, MI 49503"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3697-7767","authenticated-orcid":false,"given":"Ljubica","family":"Mihaljevi\u0107","sequence":"additional","affiliation":[{"name":"Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5421-1538","authenticated-orcid":false,"given":"Kevin Hong","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0814-161X","authenticated-orcid":false,"given":"Ami","family":"Tamhaney","sequence":"additional","affiliation":[{"name":"Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, Van Andel Institute, Grand Rapids, MI 49503"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"L\u00fc","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, Van Andel Institute, Grand Rapids, MI 49503"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3473-9818","authenticated-orcid":false,"given":"Harel","family":"Weinstein","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10065"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaozhu","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205"},{"name":"Solomon H. 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