{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T02:31:00Z","timestamp":1791945060406,"version":"4.3.4"},"reference-count":77,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2021,3,24]],"date-time":"2021-03-24T00:00:00Z","timestamp":1616544000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2021,6]]},"abstract":"<jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      <jats:list list-type=\"bullet\">\n                        <jats:list-item>\n                          <jats:p>\n                            We report a novel method for the transient expression of SARS\u2010CoV\u20102 envelope (E) protein in intracellular organelles and the plasma membrane of mammalian cells and\n                            <jats:italic>Xenopus<\/jats:italic>\n                            oocytes.\n                          <\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>Intracellular expression of SARS\u2010CoV\u20102 E protein increases intra\u2010Golgi pH.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>By targeting the SARS\u2010CoV\u20102 E protein to the plasma membrane, we show that it forms a cation channel, viroporin, that is modulated by changes of pH.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>This method for studying the activity of viroporins may facilitate screening for new antiviral drugs to identify novel treatments for COVID\u201019.<\/jats:p>\n                        <\/jats:list-item>\n                      <\/jats:list>\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      The envelope (E) protein of coronaviruses such as SARS\u2010CoV\u20101 is proposed to form an ion channel or viroporin that participates in viral propagation and pathogenesis. Here we developed a technique to study the E protein of SARS\u2010CoV\u20102 in mammalian cells by directed targeting using a carboxyl\u2010terminal fluorescent protein tag, mKate2. The wild\u2010type SARS\u2010CoV\u20102 E protein can be trafficked to intracellular organelles, notably the endoplasmic reticulum\u2013Golgi intermediate complex, where its expression increases pH inside the organelle. We also succeeded in targeting SARS\u2010CoV\u20102 E to the plasma membrane, which enabled biophysical analysis using whole\u2010cell patch clamp recording in a mammalian cell line, HEK 293 cells, and two\u2010electrode voltage clamp electrophysiology in\n                      <jats:italic>Xenopus<\/jats:italic>\n                      oocytes. The results suggest that the E protein forms an ion channel that is permeable to monovalent cations such as Na\n                      <jats:sup>+<\/jats:sup>\n                      , Cs\n                      <jats:sup>+<\/jats:sup>\n                      and K\n                      <jats:sup>+<\/jats:sup>\n                      . The E current is nearly time\u2010 and voltage\u2010independent when E protein is expressed in mammalian cells, and is modulated by changes of pH. At pH 6.0 and 7.4, the E protein current is activated, whereas at pH 8.0 and 9.0, the amplitude of E protein current is reduced, and in oocytes the inward E current fades at pH 9 in a time\u2010 and voltage\u2010dependent manner. Using this directed targeting method and electrophysiological recordings, potential inhibitors of the E protein can be screened and subsequently investigated for antiviral activity against SARS\u2010CoV\u20102\n                      <jats:italic>in vitro<\/jats:italic>\n                      and possible efficacy in treating COVID\u201019.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1113\/jp281037","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T04:32:47Z","timestamp":1615523567000},"page":"2851-2868","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":80,"title":["The envelope protein of SARS\u2010CoV\u20102 increases intra\u2010Golgi pH and forms a cation channel that is regulated by pH"],"prefix":"10.1113","volume":"599","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9609-9102","authenticated-orcid":false,"given":"David","family":"Cabrera\u2010Garcia","sequence":"first","affiliation":[{"name":"Department of Anesthesiology Columbia University  New York NY USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1825-7130","authenticated-orcid":false,"given":"Ramsey","family":"Bekdash","sequence":"additional","affiliation":[{"name":"Department of Molecular Pharmacology and Therapeutics Columbia University  New York NY USA"},{"name":"Columbia Stem Cell Initiative, Department of Rehabilitation and Regenerative Medicine Columbia University  New York NY USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geoffrey W.","family":"Abbott","sequence":"additional","affiliation":[{"name":"Bioelectricity Laboratory, Department of Physiology and Biophysics, School of Medicine University of California  Irvine CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masayuki","family":"Yazawa","sequence":"additional","affiliation":[{"name":"Department of Molecular Pharmacology and Therapeutics Columbia University  New York NY USA"},{"name":"Columbia Stem Cell Initiative, Department of Rehabilitation and Regenerative Medicine Columbia University  New York NY USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neil L.","family":"Harrison","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology Columbia University  New York NY USA"},{"name":"Department of Molecular Pharmacology and Therapeutics Columbia University  New York NY 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