Expression Cloning of a Rat Liver Na+-Independent Organic Anion Transporter
Using expression cloning in Xenopus laevis oocytes, we have isolated a cDNA encoding a rat liver organic anion-transporting polypeptide (oatp). The cloned oatp mediated Na+-independent uptake of sulfobromophthalein (BSP) which was Cl--dependent in the presence of bovine serum albumin (BSA) at low BS...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1994-01, Vol.91 (1), p.133-137 |
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| creator | Jacquemin, Emmanuel Hagenbuch, Bruno Stieger, Bruno Wolkoff, Allan W. Meier, Peter J. |
| description | Using expression cloning in Xenopus laevis oocytes, we have isolated a cDNA encoding a rat liver organic anion-transporting polypeptide (oatp). The cloned oatp mediated Na+-independent uptake of sulfobromophthalein (BSP) which was Cl--dependent in the presence of bovine serum albumin (BSA) at low BSP concentrations (e.g., 2 μM). Addition of increasing amounts of BSA had no effects on the maximal velocity of initial BSP uptake, but it increased the Kmvalue from 1.5 μM (no BSA) to 24 μM (BSA/BSP molar ratio, 3.7) and 35 μM (BSA/BSP ratio, 18.4). In addition to BSP, the cloned oatp also mediated Na+-independent uptake of conjugated (taurocholate) and unconjugated (cholate) bile acids. Sequence analysis of the cDNA revealed an open reading frame of 2010 nucleotides coding for a protein of 670 amino acids (calculated molecular mass, 74 kDa) with four possible N-linked glycosylation sites and 10 putative transmembrane domains. Translation experiments in vitro indicated that the transporter was indeed glycosylated and that its polypeptide backbone had an apparent molecular mass of 59 kDa. Northern blot analysis with the cloned probe revealed crossreactivity with several mRNA species from rat liver, kidney, brain, lung, skeletal muscle, and proximal colon as well as from liver tissues of mouse and rabbit, but not of skate (Raja erinacea) and human. |
| doi_str_mv | 10.1073/pnas.91.1.133 |
| format | Article |
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The cloned oatp mediated Na+-independent uptake of sulfobromophthalein (BSP) which was Cl--dependent in the presence of bovine serum albumin (BSA) at low BSP concentrations (e.g., 2 μM). Addition of increasing amounts of BSA had no effects on the maximal velocity of initial BSP uptake, but it increased the Kmvalue from 1.5 μM (no BSA) to 24 μM (BSA/BSP molar ratio, 3.7) and 35 μM (BSA/BSP ratio, 18.4). In addition to BSP, the cloned oatp also mediated Na+-independent uptake of conjugated (taurocholate) and unconjugated (cholate) bile acids. Sequence analysis of the cDNA revealed an open reading frame of 2010 nucleotides coding for a protein of 670 amino acids (calculated molecular mass, 74 kDa) with four possible N-linked glycosylation sites and 10 putative transmembrane domains. Translation experiments in vitro indicated that the transporter was indeed glycosylated and that its polypeptide backbone had an apparent molecular mass of 59 kDa. 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The cloned oatp mediated Na+-independent uptake of sulfobromophthalein (BSP) which was Cl--dependent in the presence of bovine serum albumin (BSA) at low BSP concentrations (e.g., 2 μM). Addition of increasing amounts of BSA had no effects on the maximal velocity of initial BSP uptake, but it increased the Kmvalue from 1.5 μM (no BSA) to 24 μM (BSA/BSP molar ratio, 3.7) and 35 μM (BSA/BSP ratio, 18.4). In addition to BSP, the cloned oatp also mediated Na+-independent uptake of conjugated (taurocholate) and unconjugated (cholate) bile acids. Sequence analysis of the cDNA revealed an open reading frame of 2010 nucleotides coding for a protein of 670 amino acids (calculated molecular mass, 74 kDa) with four possible N-linked glycosylation sites and 10 putative transmembrane domains. Translation experiments in vitro indicated that the transporter was indeed glycosylated and that its polypeptide backbone had an apparent molecular mass of 59 kDa. Northern blot analysis with the cloned probe revealed crossreactivity with several mRNA species from rat liver, kidney, brain, lung, skeletal muscle, and proximal colon as well as from liver tissues of mouse and rabbit, but not of skate (Raja erinacea) and human.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Anion Transport Proteins</subject><subject>Anions</subject><subject>Antiporters - genetics</subject><subject>Antiporters - metabolism</subject><subject>Base Sequence</subject><subject>Bile acids</subject><subject>Carrier Proteins - genetics</subject><subject>Cloning, Molecular</subject><subject>Complementary DNA</subject><subject>Complementary RNA</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Complementary - genetics</subject><subject>Gene Expression</subject><subject>Gluconates</subject><subject>Hepatocytes</subject><subject>Liver</subject><subject>Liver - metabolism</subject><subject>Membrane Glycoproteins - genetics</subject><subject>Messenger RNA</subject><subject>Molecular Sequence Data</subject><subject>Nucleotides</subject><subject>Oocytes</subject><subject>Rats</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Rodents</subject><subject>Tissue Distribution</subject><subject>Xenopus laevis</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcFrFDEUxoModa0evSkMgl5k1pe8yWQCvZSl2uJiQfYesrNv1llmkzGZKfW_N0PXbe2hPEgO3-97-cLH2FsOcw4Kv_TOxrnm8zSIz9iMg-Z5WWh4zmYAQuVVIYqX7FWMOwDQsoITdlIJVaHEGft-cdsHirH1Llt03rVum_kms9lPO2TL9oZC9sN-zq_chnpKhxuy67C1rq2zczeZVsG62PswUHjNXjS2i_TmcJ-y1deL1eIyX15_u1qcL_NaFnLI9VrwkhpSUoPVa66EkpWuFJGqCkucsFENqlLzNdaoS4ANoKySSYoGAU_Z2d3aflzvaVOnTMF2pg_t3oY_xtvW_K-49pfZ-htTCA2T_dPBHvzvkeJg9m2sqeusIz9Go0pEgaJK4IdH4M6PwaWfGQEchSyKCcrvoDr4GAM1xxwczNSPmfoxmps0iIl__zD8kT4UkvR3B32y_VMf2D8-IZtm7LqBbof7Nbs4-HAEBZao0it_Aa3CqyE</recordid><startdate>19940104</startdate><enddate>19940104</enddate><creator>Jacquemin, Emmanuel</creator><creator>Hagenbuch, Bruno</creator><creator>Stieger, Bruno</creator><creator>Wolkoff, Allan W.</creator><creator>Meier, Peter J.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19940104</creationdate><title>Expression Cloning of a Rat Liver Na+-Independent Organic Anion Transporter</title><author>Jacquemin, Emmanuel ; Hagenbuch, Bruno ; Stieger, Bruno ; Wolkoff, Allan W. ; Meier, Peter J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c545t-9b216efe7590a9b172758987ee784ae1e3f7f37691b3c39600d035821652f303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Anion Transport Proteins</topic><topic>Anions</topic><topic>Antiporters - genetics</topic><topic>Antiporters - metabolism</topic><topic>Base Sequence</topic><topic>Bile acids</topic><topic>Carrier Proteins - genetics</topic><topic>Cloning, Molecular</topic><topic>Complementary DNA</topic><topic>Complementary RNA</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Complementary - genetics</topic><topic>Gene Expression</topic><topic>Gluconates</topic><topic>Hepatocytes</topic><topic>Liver</topic><topic>Liver - metabolism</topic><topic>Membrane Glycoproteins - genetics</topic><topic>Messenger RNA</topic><topic>Molecular Sequence Data</topic><topic>Nucleotides</topic><topic>Oocytes</topic><topic>Rats</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Rodents</topic><topic>Tissue Distribution</topic><topic>Xenopus laevis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jacquemin, Emmanuel</creatorcontrib><creatorcontrib>Hagenbuch, Bruno</creatorcontrib><creatorcontrib>Stieger, Bruno</creatorcontrib><creatorcontrib>Wolkoff, Allan W.</creatorcontrib><creatorcontrib>Meier, Peter J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jacquemin, Emmanuel</au><au>Hagenbuch, Bruno</au><au>Stieger, Bruno</au><au>Wolkoff, Allan W.</au><au>Meier, Peter J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression Cloning of a Rat Liver Na+-Independent Organic Anion Transporter</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1994-01-04</date><risdate>1994</risdate><volume>91</volume><issue>1</issue><spage>133</spage><epage>137</epage><pages>133-137</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Using expression cloning in Xenopus laevis oocytes, we have isolated a cDNA encoding a rat liver organic anion-transporting polypeptide (oatp). 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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1994-01, Vol.91 (1), p.133-137 |
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| source | MEDLINE; Jstor Complete Legacy; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence Animals Anion Transport Proteins Anions Antiporters - genetics Antiporters - metabolism Base Sequence Bile acids Carrier Proteins - genetics Cloning, Molecular Complementary DNA Complementary RNA Deoxyribonucleic acid DNA DNA, Complementary - genetics Gene Expression Gluconates Hepatocytes Liver Liver - metabolism Membrane Glycoproteins - genetics Messenger RNA Molecular Sequence Data Nucleotides Oocytes Rats RNA, Messenger - genetics RNA, Messenger - metabolism Rodents Tissue Distribution Xenopus laevis |
| title | Expression Cloning of a Rat Liver Na+-Independent Organic Anion Transporter |
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