{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T15:41:36Z","timestamp":1759333296099,"version":"3.30.0"},"reference-count":68,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"6","license":[{"start":{"date-parts":[[2000,6,1]],"date-time":"2000-06-01T00:00:00Z","timestamp":959817600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"},{"start":{"date-parts":[[2003,12,30]],"date-time":"2003-12-30T00:00:00Z","timestamp":1072742400000},"content-version":"vor","delay-in-days":1307,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2000,6]]},"abstract":"<jats:sec>\n            <jats:title\/>\n            <jats:p>We have cloned two complementary DNAs (cDNAs), RL-Mtx-1 and RL-Mtx-2, corresponding to the bile acid- sensitive methotrexate carrier from rat liver by direct full-length rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR) using degenerated primers that were deduced from published sequences of tumor cell methotrexate transporters. When expressed in <jats:italic toggle=\"yes\">Xenopus laevis<\/jats:italic> oocytes and cosM6 cells, both clones mediate methotrexate and bumetanide transport. RL-Mtx-1 consists of 2,445 bp with an open reading frame of 1,536 bp. The corresponding protein with 512 amino acids has a molecular weight of 58 kd. RL-Mtx-2 (2,654 bp) differs by an additional insert of 203 bp. This insert is located in frame at position 1,196 of the RL-Mtx-1 and contains the typical splice junction sites at the 5\u2032 and 3\u2032 end, indicating that the RL-Mtx-2 messenger RNA (mRNA) is generated by alternative splicing. The insert contains a stop codon that shortens the RL-Mtx-2 protein to 330 amino acids (38 kd). Both cDNAs contain the binding site sequence for the dioxin\/nuclear translocator responsive element (Ah\/Arnt-receptor) in conjunction with a barbiturate recognition sequence (Barbie box). Preliminary results show that the Barbie box acts as a negative regulatory element. The two liver cDNA clones show homologies to the published sequences of folate and the reduced folate carriers, but no homology is found to the transport systems for organic anions like the Ntcp1, oatp1, OAT-K1, and OAT1. Expression of the mRNA for the methotrexate carrier is found in liver, kidney, heart, brain, spleen, lung, and skeletal muscle, but not in the testis as revealed by Northern blot analysis. The highest abundance of the mRNA is found in the kidney.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1053\/jhep.2000.7478","type":"journal-article","created":{"date-parts":[[2004,3,19]],"date-time":"2004-03-19T18:37:43Z","timestamp":1079721463000},"page":"1296-1304","update-policy":"http:\/\/dx.doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":18,"title":["Cloning and functional characterization of the bile acid-sensitive methotrexate carrier from rat liver cells"],"prefix":"10.1097","volume":"31","author":[{"given":"Walther","family":"Honscha","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kerstin U.","family":"D\u00f6tsch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nadine","family":"Thomsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ernst","family":"Petzinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"276","reference":[{"key":"10.1053\/jhep.2000.7478-BIB1|cit1","doi-asserted-by":"crossref","first-page":"357","DOI":"10.3727\/095535491820873687","article-title":"Folate transport and the modulation of antifolate sensitivity in a methotrexate-resistant human breast cancer cell line","volume":"3","author":"Dixon","year":"1991","journal-title":"Cancer Commun"},{"key":"10.1053\/jhep.2000.7478-BIB2|cit2","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1007\/BF00689044","article-title":"Pharmacokinetics of different doses of methotrexate at steady state by in situ microdialysis in a rat model","volume":"36","author":"Ekstr\u00f8m","year":"1995","journal-title":"Cancer Chemother Pharmacol"},{"key":"10.1053\/jhep.2000.7478-BIB3|cit3","doi-asserted-by":"crossref","first-page":"14911","DOI":"10.1016\/S0021-9258(18)95679-6","article-title":"Studies of the role of a particulate folate-binding protein in the uptake of 5-methyltetrahydrofolate by cultured human KB cells","volume":"260","author":"Antony","year":"1985","journal-title":"J Biol Chem"},{"key":"10.1053\/jhep.2000.7478-BIB4|cit4","doi-asserted-by":"crossref","first-page":"15416","DOI":"10.1016\/S0021-9258(18)66727-4","article-title":"The isolation, characterization, and comparison of the membrane-associated and soluble folate-binding proteins from human KB cells","volume":"261","author":"Elwood","year":"1986","journal-title":"J Biol Chem"},{"key":"10.1053\/jhep.2000.7478-BIB5|cit5","doi-asserted-by":"crossref","first-page":"15625","DOI":"10.1016\/S0021-9258(18)66761-4","article-title":"The interrelationship of the soluble and membrane-associated folate-binding proteins in human KB cells","volume":"261","author":"Kane","year":"1986","journal-title":"J Biol Chem"},{"key":"10.1053\/jhep.2000.7478-BIB6|cit6","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/S0021-9258(17)42427-6","article-title":"The influence of extracellular folate concentration on methotrexate uptake by human KB cells. 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