{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T09:32:14Z","timestamp":1775554334394,"version":"3.50.1"},"reference-count":48,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T00:00:00Z","timestamp":1753142400000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal Cellular Physiology"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    SARS\u2010CoV\u20102 binds to its obligatory receptor, angiotensin\u2010converting enzyme 2 (ACE2) and capitalizes on decreasing endosomal acidity and cathepsin\u2010mediated spike protein cleavage to enter cells. Endosomal acidification is driven by V\u2010ATPase which pumps protons (H\n                    <jats:sup>+<\/jats:sup>\n                    ) into the lumen. The driving force for H\n                    <jats:sup>+<\/jats:sup>\n                    is maintained by the import of chloride (Cl\n                    <jats:sup>\u2212<\/jats:sup>\n                    ) which is mediated by intracellular CLC transporters. We have recently identified the Proton\u2010Activated Chloride (PAC) channel as a negative regulator of endosomal acidification. PAC responds to low pH and releases Cl\n                    <jats:sup>\u2212<\/jats:sup>\n                    from the lumen to prevent endosomal hyperacidification. However, its role in SARS\u2010CoV\u20102 viral entry remains unexplored. Here, we show that overexpressing the PAC channel in ACE2 expressing HEK 293T cells markedly inhibited SARS\u2010CoV\u20102 spike\u2010mediated viral entry. Several lines of evidence suggest that this effect was due to the suppression of the endosomal entry pathway. First, the abilities of PAC to regulate endosomal acidification and inhibit pseudoviral entry were both dependent on its endosomal localization and channel activity. Second, the inhibitory effect on viral entry was similar to the suppression mediated by E64\u2010d, a cathepsin inhibitor, while no major additive effect for both treatments was observed. Third, this inhibition was also attenuated in cells expressing TMPRSS2, which provides an alternative entry pathway through the cell surface. Importantly, PAC overexpression also inhibited the number and size of plaques formed by two live SARS\u2010CoV\u20102 isolates (B.1 and Omicron XBB.1.16) in Vero E6 cells. Altogether, our data indicates that PAC plays a vital role in inhibiting SARS\u2010CoV\u20102 viral entry and identifies this endosomal channel as a potential novel target against the infection of SARS\u2010CoV\u20102 and other viruses, which rely on the endosomal pathway.\n                  <\/jats:p>","DOI":"10.1002\/jcp.70063","type":"journal-article","created":{"date-parts":[[2025,7,23]],"date-time":"2025-07-23T06:15:33Z","timestamp":1753251333000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["The Proton\u2010Activated Chloride Channel Inhibits SARS\u2010CoV\u20102 Spike Protein\u2010Mediated Viral Entry Through the Endosomal Pathway"],"prefix":"10.1002","volume":"240","author":[{"given":"Nicholas","family":"Koylass","sequence":"first","affiliation":[{"name":"Department of Physiology Johns Hopkins University School of Medicine Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaiprasath","family":"Sachithanandham","sequence":"additional","affiliation":[{"name":"W. Harry Feinstone Department of Molecular Microbiology and Immunology Johns Hopkins University Bloomberg School of Public Health Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James","family":"Osei\u2010Owusu","sequence":"additional","affiliation":[{"name":"Department of Physiology Johns Hopkins University School of Medicine Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin Hong","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Physiology Johns Hopkins University School of Medicine Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henry Yi","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Physiology Johns Hopkins University School of Medicine Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew","family":"Pekosz","sequence":"additional","affiliation":[{"name":"W. Harry Feinstone Department of Molecular Microbiology and Immunology Johns Hopkins University Bloomberg School of Public Health Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9122-6077","authenticated-orcid":false,"given":"Zhaozhu","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Physiology Johns Hopkins University School of Medicine Baltimore MD USA"},{"name":"Solomon H. Snyder Department of Neuroscience Johns Hopkins University School of Medicine Baltimore MD USA"},{"name":"Department of Neurosurgery Johns Hopkins University School of Medicine Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,7,22]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13075-021-02700-x"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100306"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41588-022-01131-x"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4073"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21249739"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm2820"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/v12080844"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1080\/15563650500514558"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2025.115302"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1101\/2025.02.27.640612"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmed.2009.05.003"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-52773-w"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2024.108144"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41401-020-0485-4"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.201210930"},{"issue":"13","key":"e_1_2_8_17_1","first-page":"6100","article-title":"Essential Role of TMPRSS2 in SARS\u2010CoV\u20102 Infection In Murine Airways","volume":"13","author":"Iwata\u2010Yoshikawa N.","year":"2022","journal-title":"Nature Communications 2022 13"},{"issue":"23","key":"e_1_2_8_18_1","first-page":"3","article-title":"Mechanisms of SARS\u2010CoV\u20102 Entry Into Cells","volume":"1","author":"Jackson C. 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