{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T18:30:22Z","timestamp":1793039422196,"version":"4.5.0"},"reference-count":39,"publisher":"Rockefeller University Press","issue":"1","license":[{"start":{"date-parts":[[2020,11,19]],"date-time":"2020-11-19T00:00:00Z","timestamp":1605744000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/www.rupress.org\/terms\/"},{"start":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T00:00:00Z","timestamp":1625443200000},"content-version":"vor","delay-in-days":228,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-sa\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000674","name":"Physiological Society","doi-asserted-by":"publisher","award":["444"],"award-info":[{"award-number":["444"]}],"id":[{"id":"10.13039\/501100000674","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009187","name":"Medical Research Fund","doi-asserted-by":"publisher","award":["23\/2016"],"award-info":[{"award-number":["23\/2016"]}],"id":[{"id":"10.13039\/501100009187","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007076","name":"Fondazione AIRC per la Ricerca sul Cancro","doi-asserted-by":"publisher","award":["IG 21558"],"award-info":[{"award-number":["IG 21558"]}],"id":[{"id":"10.13039\/501100007076","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003407","name":"Italian Research Ministry","doi-asserted-by":"publisher","award":["PRIN 20174TB8KW"],"award-info":[{"award-number":["PRIN 20174TB8KW"]}],"id":[{"id":"10.13039\/501100003407","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,1,4]]},"abstract":"<jats:p>ClC-7 is a lysosomal 2 Cl\u2212\/1 H+ antiporter of the CLC protein family, which comprises Cl\u2212 channels and other Cl\u2212\/H+ antiporters. Mutations in ClC-7 and its associated \u03b2 subunit Ostm1 lead to osteopetrosis and lysosomal storage disease in humans and mice. Previous studies on other mammalian CLC transporters showed that mutations of a conserved, intracellularly located glutamate residue, the so-called proton glutamate, abolish steady-state transport activity but increase transient capacitive currents associated with partial reactions of the transport cycle. In contrast, we observed large, transient capacitive currents for the wild-type ClC-7, which depend on external pH and internal, but not external, Cl\u2212. Very similar transient currents were observed for the E312A mutant of the proton glutamate. Interestingly, and unlike in other mammalian CLC transporters investigated so far, the E312A mutation strongly reduces, but does not abolish, stationary transport currents, potentially explaining the intermediate phenotype observed in the E312A mouse line.<\/jats:p>","DOI":"10.1085\/jgp.202012583","type":"journal-article","created":{"date-parts":[[2020,11,19]],"date-time":"2020-11-19T16:59:07Z","timestamp":1605805147000},"update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":15,"title":["Large transient capacitive currents in wild-type lysosomal Cl\u2212\/H+ antiporter ClC-7 and residual transport activity in the proton glutamate mutant E312A"],"prefix":"10.1085","volume":"153","author":[{"given":"Michael","family":"Pusch","sequence":"first","affiliation":[{"name":"Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5567-8300","authenticated-orcid":true,"given":"Giovanni","family":"Zifarelli","sequence":"additional","affiliation":[{"name":"Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2020,11,19]]},"reference":[{"key":"2023072706105178400_bib1","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1085\/jgp.200308834","article-title":"Conformational changes in the pore of CLC-0","volume":"122","author":"Accardi","year":"2003","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib2","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1085\/jgp.200308935","article-title":"Ionic currents mediated by a prokaryotic homologue of CLC Cl- channels","volume":"123","author":"Accardi","year":"2004","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib3","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1085\/jgp.200509417","article-title":"Separate ion pathways in a Cl-\/H+ exchanger","volume":"126","author":"Accardi","year":"2005","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib4","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1038\/nrm2376","article-title":"How membrane proteins sense voltage","volume":"9","author":"Bezanilla","year":"2008","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2023072706105178400_bib5","doi-asserted-by":"publisher","first-page":"5879","DOI":"10.1113\/jphysiol.2013.260240","article-title":"A single point mutation reveals gating of the human ClC-5 Cl-\/H+ antiporter","volume":"591","author":"De Stefano","year":"2013","journal-title":"J. Physiol."},{"key":"2023072706105178400_bib6","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1038\/415287a","article-title":"X-ray structure of a ClC chloride channel at 3.0 A reveals the molecular basis of anion selectivity","volume":"415","author":"Dutzler","year":"2002","journal-title":"Nature."},{"key":"2023072706105178400_bib7","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1126\/science.1082708","article-title":"Gating the selectivity filter in ClC chloride channels","volume":"300","author":"Dutzler","year":"2003","journal-title":"Science."},{"key":"2023072706105178400_bib8","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1126\/science.1195230","article-title":"Structure of a eukaryotic CLC transporter defines an intermediate state in the transport cycle","volume":"330","author":"Feng","year":"2010","journal-title":"Science."},{"key":"2023072706105178400_bib9","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1038\/nature06907","article-title":"The Cl-\/H+ antiporter ClC-7 is the primary chloride permeation pathway in lysosomes","volume":"453","author":"Graves","year":"2008","journal-title":"Nature."},{"key":"2023072706105178400_bib10","doi-asserted-by":"publisher","first-page":"8101","DOI":"10.1074\/jbc.M111.298265","article-title":"Glutamate 268 regulates transport probability of the anion\/proton exchanger ClC-5","volume":"287","author":"Grieschat","year":"2012","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib11","doi-asserted-by":"publisher","first-page":"8075","DOI":"10.1073\/pnas.95.14.8075","article-title":"ClC-5, the chloride channel mutated in Dent\u2019s disease, colocalizes with the proton pump in endocytotically active kidney cells","volume":"95","author":"G\u00fcnther","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072706105178400_bib12","doi-asserted-by":"publisher","first-page":"994","DOI":"10.1021\/cn400032z","article-title":"ClC-3 is an intracellular chloride\/proton exchanger with large voltage-dependent nonlinear capacitance","volume":"4","author":"Guzman","year":"2013","journal-title":"ACS Chem. Neurosci."},{"key":"2023072706105178400_bib13","doi-asserted-by":"publisher","first-page":"40169","DOI":"10.1074\/jbc.M307890200","article-title":"Cofilin interacts with ClC-5 and regulates albumin uptake in proximal tubule cell lines","volume":"278","author":"Hryciw","year":"2003","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib14","doi-asserted-by":"publisher","first-page":"1493","DOI":"10.1152\/physrev.00047.2017","article-title":"CLC Chloride Channels and Transporters: Structure, Function, Physiology, and Disease","volume":"98","author":"Jentsch","year":"2018","journal-title":"Physiol. Rev."},{"key":"2023072706105178400_bib15","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1038\/sj.emboj.7600576","article-title":"Loss of the chloride channel ClC-7 leads to lysosomal storage disease and neurodegeneration","volume":"24","author":"Kasper","year":"2005","journal-title":"EMBO J."},{"key":"2023072706105178400_bib16","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1016\/S0092-8674(01)00206-9","article-title":"Loss of the ClC-7 chloride channel leads to osteopetrosis in mice and man","volume":"104","author":"Kornak","year":"2001","journal-title":"Cell."},{"key":"2023072706105178400_bib17","doi-asserted-by":"publisher","first-page":"220","DOI":"10.1038\/nature04535","article-title":"ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function","volume":"440","author":"Lange","year":"2006","journal-title":"Nature."},{"key":"2023072706105178400_bib18","doi-asserted-by":"publisher","first-page":"2140","DOI":"10.1038\/emboj.2011.137","article-title":"ClC-7 is a slowly voltage-gated 2Cl(-)\/1H(+)-exchanger and requires Ostm1 for transport activity","volume":"30","author":"Leisle","year":"2011","journal-title":"EMBO J."},{"key":"2023072706105178400_bib19","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1085\/jgp.200810112","article-title":"Intracellular proton-transfer mutants in a CLC Cl-\/H+ exchanger","volume":"133","author":"Lim","year":"2009","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib20","doi-asserted-by":"publisher","first-page":"28611","DOI":"10.1074\/jbc.M113.509364","article-title":"Common gating of both CLC transporter subunits underlies voltage-dependent activation of the 2Cl-\/1H+ exchanger ClC-7\/Ostm1","volume":"288","author":"Ludwig","year":"2013","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib21","doi-asserted-by":"publisher","first-page":"401","DOI":"10.1098\/rstb.1982.0140","article-title":"Open-state substructure of single chloride channels from Torpedo electroplax","volume":"299","author":"Miller","year":"1982","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"2023072706105178400_bib22","doi-asserted-by":"publisher","first-page":"21689","DOI":"10.1074\/jbc.M110.125971","article-title":"The late endosomal ClC-6 mediates proton\/chloride countertransport in heterologous plasma membrane expression","volume":"285","author":"Neagoe","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib23","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.36629","article-title":"Structure of the CLC-1 chloride channel from Homo sapiens","volume":"7","author":"Park","year":"2018","journal-title":"eLife."},{"key":"2023072706105178400_bib24","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1038\/nature20812","article-title":"Structure of a CLC chloride ion channel by cryo-electron microscopy","volume":"541","author":"Park","year":"2017","journal-title":"Nature."},{"key":"2023072706105178400_bib25","doi-asserted-by":"publisher","first-page":"420","DOI":"10.1038\/nature03720","article-title":"Chloride\/proton antiporter activity of mammalian CLC proteins ClC-4 and ClC-5","volume":"436","author":"Picollo","year":"2005","journal-title":"Nature."},{"key":"2023072706105178400_bib26","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1085\/jgp.113.3.457","article-title":"The muscle chloride channel ClC-1 has a double-barreled appearance that is differentially affected in dominant and recessive myotonia","volume":"113","author":"Saviane","year":"1999","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib27","doi-asserted-by":"publisher","first-page":"424","DOI":"10.1038\/nature03860","article-title":"Voltage-dependent electrogenic chloride\/proton exchange by endosomal CLC proteins","volume":"436","author":"Scheel","year":"2005","journal-title":"Nature."},{"key":"2023072706105178400_bib28","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.59555","article-title":"Cryo-EM structure of the lysosomal chloride-proton exchanger CLC-7 in complex with OSTM1","volume":"9","author":"Schrecker","year":"2020","journal-title":"eLife."},{"key":"2023072706105178400_bib29","doi-asserted-by":"publisher","first-page":"3696","DOI":"10.1096\/fj.09-150649","article-title":"Voltage-dependent charge movement associated with activation of the CLC-5 2Cl-\/1H+ exchanger","volume":"24","author":"Smith","year":"2010","journal-title":"FASEB J."},{"key":"2023072706105178400_bib30","doi-asserted-by":"publisher","first-page":"34537","DOI":"10.1074\/jbc.M110.162545","article-title":"Sorting motifs of the endosomal\/lysosomal CLC chloride transporters","volume":"285","author":"Stauber","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib31","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1085\/jgp.200709756","article-title":"Uncoupling and turnover in a Cl-\/H+ exchange transporter","volume":"129","author":"Walden","year":"2007","journal-title":"J. Gen. Physiol."},{"key":"2023072706105178400_bib32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3000218","article-title":"Structure of the human ClC-1 chloride channel","volume":"17","author":"Wang","year":"2019","journal-title":"PLoS Biol."},{"key":"2023072706105178400_bib33","doi-asserted-by":"publisher","first-page":"1401","DOI":"10.1126\/science.1188072","article-title":"Lysosomal pathology and osteopetrosis upon loss of H+-driven lysosomal Cl- accumulation","volume":"328","author":"Weinert","year":"2010","journal-title":"Science."},{"key":"2023072706105178400_bib34","doi-asserted-by":"publisher","first-page":"784","DOI":"10.15252\/embr.201438553","article-title":"Transport activity and presence of ClC-7\/Ostm1 complex account for different cellular functions","volume":"15","author":"Weinert","year":"2014","journal-title":"EMBO Rep."},{"key":"2023072706105178400_bib35","doi-asserted-by":"publisher","first-page":"1231","DOI":"10.1038\/srep01231","article-title":"An optical assay of the transport activity of ClC-7","volume":"3","author":"Zanardi","year":"2013","journal-title":"Sci. Rep."},{"key":"2023072706105178400_bib36","doi-asserted-by":"publisher","first-page":"4219","DOI":"10.1074\/jbc.M708368200","article-title":"Determinants of anion-proton coupling in mammalian endosomal CLC proteins","volume":"283","author":"Zdebik","year":"2008","journal-title":"J. Biol. Chem."},{"key":"2023072706105178400_bib37","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abb4747","article-title":"Molecular insights into the human CLC-7\/Ostm1 transporter","volume":"6","author":"Zhang","year":"2020","journal-title":"Sci. Adv."},{"key":"2023072706105178400_bib38","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1007\/112_2006_0605","article-title":"CLC chloride channels and transporters: a biophysical and physiological perspective","volume":"158","author":"Zifarelli","year":"2007","journal-title":"Rev. Physiol. Biochem. Pharmacol."},{"key":"2023072706105178400_bib39","doi-asserted-by":"publisher","first-page":"2060","DOI":"10.1016\/j.bpj.2012.03.067","article-title":"On the mechanism of gating charge movement of ClC-5, a human Cl-\/H+ antiporter","volume":"102","author":"Zifarelli","year":"2012","journal-title":"Biophys. J."}],"container-title":["Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/doi\/10.1085\/jgp.202012583\/1801712\/jgp_202012583.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/doi\/10.1085\/jgp.202012583\/1801712\/jgp_202012583.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,27]],"date-time":"2023-07-27T06:11:41Z","timestamp":1690438301000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/153\/1\/e202012583\/211547\/Large-transient-capacitive-currents-in-wild-type"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,19]]},"references-count":39,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,1,4]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.202012583","relation":{},"ISSN":["0022-1295","1540-7748"],"issn-type":[{"value":"0022-1295","type":"print"},{"value":"1540-7748","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,19]]},"article-number":"e202012583"}}