{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T21:34:28Z","timestamp":1770500068878,"version":"3.49.0"},"reference-count":43,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"5","license":[{"start":{"date-parts":[[2015,9,1]],"date-time":"2015-09-01T00:00:00Z","timestamp":1441065600000},"content-version":"tdm","delay-in-days":6148,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1998,11]]},"abstract":"<jats:sec>\n            <jats:title\/>\n            <jats:p>The polarized rat hepatoma\/human fibroblast hybrid cell line, WIF\u2013B, forms apical vacuoles into which cholephilic substances are secreted. We studied expression, localization, and function of the apical conjugate export pump, Mrp2, in WIF\u2013B cells. Mrp2, the apical isoform of the multidrug resistance protein, alternatively termed canalicular Mrp (cMrp) or canalicular multispecific organic anion transporter (cMoat), is a 190\u2013kd membrane glycoprotein mediating adenosine triphosphate (ATP)\u2013dependent transport of glucuronides, glutathione <jats:italic toggle=\"yes\">S<\/jats:italic>\u2013conjugates, and other amphiphilic anions across the hepatocyte canalicular membrane into bile. Expression of the rat <jats:italic toggle=\"yes\">mrp2<\/jats:italic> gene in WIF\u2013B cells was shown by reverse\u2013transcription polymerase chain reaction (PCR), followed by sequencing of the amplified 789\u2013bp fragment. Immunoblotting, using antibodies reacting with the amino\u2013terminal or with the carboxyl\u2013terminal sequence of rat Mrp2, detected the 190\u2013kd glycoprotein in WIF\u2013B cell homogenates. Immunofluorescence microscopy localized Mrp2 to the apical membrane domain. Preloading of WIF\u2013B cells with a membrane\u2013permeable ester of the calcium\u2013dependent fluorescent indicator, Fluo\u20133, was followed by Mrp2\u2013mediated secretion of the amphiphilic anion, Fluo\u20133, into the apical vacuoles. This transport was potently inhibited by cyclosporin A added to the culture medium. Direct measurements of ATP\u2013dependent transport into Mrp2\u2013containing plasma membrane vesicles in comparison with Mrp2\u2013deficient vesicles established that Fluo\u20133 is transported by Mrp2 with a <jats:italic toggle=\"yes\">K<\/jats:italic>\n                                 <jats:sub>m<\/jats:sub> value of 3.7 \u03bcmol\/L. Our results indicate that the polarized WIF\u2013B cells express the rat ortholog of the apical conjugate-transporting ATPase, Mrp2. The function of Mrp2 as well as the action of inhibitors can thus be analyzed by use of the fluorescent amphiphilic anion, Fluo\u20133.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1002\/hep.510280523","type":"journal-article","created":{"date-parts":[[2004,7,27]],"date-time":"2004-07-27T23:30:02Z","timestamp":1090971002000},"page":"1332-1340","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":79,"title":["Expression of the Apical Conjugate Export Pump, Mrp2, in the Polarized Hepatoma Cell Line, Wif\u2013B"],"prefix":"10.1097","volume":"28","author":[{"given":"Anne T.","family":"Nies","sequence":"first","affiliation":[]},{"given":"Tobias","family":"Cantz","sequence":"additional","affiliation":[]},{"given":"Manuela","family":"Brom","sequence":"additional","affiliation":[]},{"given":"Inka","family":"Leier","sequence":"additional","affiliation":[]},{"given":"Dietrich","family":"Keppler","sequence":"additional","affiliation":[]}],"member":"276","reference":[{"key":"BIB1","doi-asserted-by":"crossref","first-page":"1667","DOI":"10.1002\/hep.510260641","volume":"26","author":"Meier","year":"1997","journal-title":"Hepatology"},{"key":"BIB2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1096\/fasebj.11.1.9034161","volume":"11","author":"Keppler","year":"1997","journal-title":"FASEB J"},{"key":"BIB3","doi-asserted-by":"crossref","first-page":"15091","DOI":"10.1074\/jbc.271.25.15091","volume":"271","author":"B\u00fcchler","year":"1996","journal-title":"J Biol Chem"},{"key":"BIB4","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1126\/science.271.5252.1126","volume":"271","author":"Paulusma","year":"1996","journal-title":"Science"},{"key":"BIB5","first-page":"4124","volume":"56","author":"Taniguchi","year":"1996","journal-title":"Cancer Res"},{"key":"BIB6","first-page":"g16","volume":"272","author":"Ito","year":"1997","journal-title":"Am J Physiol"},{"key":"BIB7","first-page":"55","volume-title":"Progress in Liver Diseases.","author":"Keppler","year":"1996","unstructured":"The canalicular conjugate export pump encoded by the cmrp\/cmoat gene. 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