Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol
A cDNA clone encoding the major intestinal cytosolic 14-kDa bile acid-binding protein (14-kDa I-BABP) was isolated from a rat ileal lambda gt22A library following immunoscreening using a monospecific antiserum raised against a 14-kDa polypeptide found in the rat ileal cytosol. One done of 516 bp enc...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1994-05, Vol.91 (11), p.4741-4745 |
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| creator | Gong, Yong-Zhong Everett, Eric T. Schwartz, David A. Norris, James S. Wilson, Frederick A. |
| description | A cDNA clone encoding the major intestinal cytosolic 14-kDa bile acid-binding protein (14-kDa I-BABP) was isolated from a rat ileal lambda gt22A library following immunoscreening using a monospecific antiserum raised against a 14-kDa polypeptide found in the rat ileal cytosol. One done of 516 bp encoded a 128-amino acid protein with a predicted molecular mass of 14,544 Da. The deduced amino acid sequence of 14-kDa I-BABP showed 100% homology to rat intestinal 15-kDa protein (I-15P) and 72% homology to porcine 15-kDa gastrotropin, whereas comparison of I-BABP to rat 14-kDa fatty acid-binding proteins of liver, intestine, and heart revealed homologies of 44%, 25%, and 28%, respectively. Northern blot analysis revealed a single transcript of approximately 0.5 kb in ileum and ovary; however, the abundance of I-BABP mRNA was much greater in ileum than in ovary. No transcript was seen in RNA extracted from stomach. jejunum, colon, liver, adrenal, brain, heart, kidney, or testis. Transfection of the I-BABP cDNA into COS-7 cells resulted in the expression of a 14-kDa protein that was identical to the ileal cytosolic I-BABP as determined by immunoblotting. Photoaffinity labeling of expressed 14-kDa protein was saturable with respect to increasing concentrations of 7,7-azo[(3)H]taurocholate [K(m), 83.3 micromolars; V(max), 6.7 pmol/mg per 5 min]. Taurocholate inhibited 7,7-azotaurocholate labeling by >96% with lesser inhibition by taurochenodeoxycholate (83.1%), chenodeoxycholate (74.6%), cholate (50.5%), and progesterone (38.5%), whereas oleic acid and estradiol did not inhibit binding. |
| doi_str_mv | 10.1073/pnas.91.11.4741 |
| format | Article |
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One done of 516 bp encoded a 128-amino acid protein with a predicted molecular mass of 14,544 Da. The deduced amino acid sequence of 14-kDa I-BABP showed 100% homology to rat intestinal 15-kDa protein (I-15P) and 72% homology to porcine 15-kDa gastrotropin, whereas comparison of I-BABP to rat 14-kDa fatty acid-binding proteins of liver, intestine, and heart revealed homologies of 44%, 25%, and 28%, respectively. Northern blot analysis revealed a single transcript of approximately 0.5 kb in ileum and ovary; however, the abundance of I-BABP mRNA was much greater in ileum than in ovary. No transcript was seen in RNA extracted from stomach. jejunum, colon, liver, adrenal, brain, heart, kidney, or testis. Transfection of the I-BABP cDNA into COS-7 cells resulted in the expression of a 14-kDa protein that was identical to the ileal cytosolic I-BABP as determined by immunoblotting. Photoaffinity labeling of expressed 14-kDa protein was saturable with respect to increasing concentrations of 7,7-azo[(3)H]taurocholate [K(m), 83.3 micromolars; V(max), 6.7 pmol/mg per 5 min]. Taurocholate inhibited 7,7-azotaurocholate labeling by >96% with lesser inhibition by taurochenodeoxycholate (83.1%), chenodeoxycholate (74.6%), cholate (50.5%), and progesterone (38.5%), whereas oleic acid and estradiol did not inhibit binding.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.91.11.4741</identifier><identifier>PMID: 8197128</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>acide biliaire ; acidos biliares ; Amino Acid Sequence ; Amino acids ; Animals ; arn mensajero ; arn messager ; Base Sequence ; Bile ; Bile acids ; Bile Acids and Salts - metabolism ; binding proteins ; Biochemistry ; Blotting, Northern ; Blotting, Western ; Carrier Proteins - biosynthesis ; Carrier Proteins - genetics ; Carrier Proteins - metabolism ; Cell Line ; Cells ; citoplasma ; Cloning, Molecular ; Complementary DNA ; COS cells ; cytoplasm ; cytoplasme ; Cytosol - metabolism ; Deoxyribonucleic acid ; DNA ; expresion genica ; expression des genes ; gene ; gene expression ; genes ; Gongs ; Humans ; Hydroxysteroid Dehydrogenases ; Ileum ; Ileum - metabolism ; intestin ; intestines ; intestinos ; Liver ; Membrane Glycoproteins ; messenger rna ; Molecular Sequence Data ; nucleotide sequence ; Organ Specificity - genetics ; proteinas aglutinantes ; proteine de liaison ; Proteins ; rat ; rata ; Rats ; Rats, Sprague-Dawley ; RNA ; Rodents ; secuencia nucleica ; Sequence Homology, Amino Acid ; sequence nucleique ; Transfection</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1994-05, Vol.91 (11), p.4741-4745</ispartof><rights>Copyright 1994 The National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences May 24, 1994</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c609t-8650b2462ea954446be9644c3f85e60e3c73ead1f5e3290e0b7178a2b25bcc523</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/91/11.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2364845$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2364845$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,315,728,781,785,804,886,27929,27930,53796,53798,58022,58255</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8197128$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gong, Yong-Zhong</creatorcontrib><creatorcontrib>Everett, Eric T.</creatorcontrib><creatorcontrib>Schwartz, David A.</creatorcontrib><creatorcontrib>Norris, James S.</creatorcontrib><creatorcontrib>Wilson, Frederick A.</creatorcontrib><title>Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>A cDNA clone encoding the major intestinal cytosolic 14-kDa bile acid-binding protein (14-kDa I-BABP) was isolated from a rat ileal lambda gt22A library following immunoscreening using a monospecific antiserum raised against a 14-kDa polypeptide found in the rat ileal cytosol. One done of 516 bp encoded a 128-amino acid protein with a predicted molecular mass of 14,544 Da. The deduced amino acid sequence of 14-kDa I-BABP showed 100% homology to rat intestinal 15-kDa protein (I-15P) and 72% homology to porcine 15-kDa gastrotropin, whereas comparison of I-BABP to rat 14-kDa fatty acid-binding proteins of liver, intestine, and heart revealed homologies of 44%, 25%, and 28%, respectively. Northern blot analysis revealed a single transcript of approximately 0.5 kb in ileum and ovary; however, the abundance of I-BABP mRNA was much greater in ileum than in ovary. No transcript was seen in RNA extracted from stomach. jejunum, colon, liver, adrenal, brain, heart, kidney, or testis. Transfection of the I-BABP cDNA into COS-7 cells resulted in the expression of a 14-kDa protein that was identical to the ileal cytosolic I-BABP as determined by immunoblotting. Photoaffinity labeling of expressed 14-kDa protein was saturable with respect to increasing concentrations of 7,7-azo[(3)H]taurocholate [K(m), 83.3 micromolars; V(max), 6.7 pmol/mg per 5 min]. Taurocholate inhibited 7,7-azotaurocholate labeling by >96% with lesser inhibition by taurochenodeoxycholate (83.1%), chenodeoxycholate (74.6%), cholate (50.5%), and progesterone (38.5%), whereas oleic acid and estradiol did not inhibit binding.</description><subject>acide biliaire</subject><subject>acidos biliares</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Animals</subject><subject>arn mensajero</subject><subject>arn messager</subject><subject>Base Sequence</subject><subject>Bile</subject><subject>Bile acids</subject><subject>Bile Acids and Salts - metabolism</subject><subject>binding proteins</subject><subject>Biochemistry</subject><subject>Blotting, Northern</subject><subject>Blotting, Western</subject><subject>Carrier Proteins - biosynthesis</subject><subject>Carrier Proteins - genetics</subject><subject>Carrier Proteins - metabolism</subject><subject>Cell Line</subject><subject>Cells</subject><subject>citoplasma</subject><subject>Cloning, Molecular</subject><subject>Complementary DNA</subject><subject>COS cells</subject><subject>cytoplasm</subject><subject>cytoplasme</subject><subject>Cytosol - metabolism</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>expresion genica</subject><subject>expression des genes</subject><subject>gene</subject><subject>gene expression</subject><subject>genes</subject><subject>Gongs</subject><subject>Humans</subject><subject>Hydroxysteroid Dehydrogenases</subject><subject>Ileum</subject><subject>Ileum - metabolism</subject><subject>intestin</subject><subject>intestines</subject><subject>intestinos</subject><subject>Liver</subject><subject>Membrane Glycoproteins</subject><subject>messenger rna</subject><subject>Molecular Sequence Data</subject><subject>nucleotide sequence</subject><subject>Organ Specificity - genetics</subject><subject>proteinas aglutinantes</subject><subject>proteine de liaison</subject><subject>Proteins</subject><subject>rat</subject><subject>rata</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>RNA</subject><subject>Rodents</subject><subject>secuencia nucleica</subject><subject>Sequence Homology, Amino Acid</subject><subject>sequence nucleique</subject><subject>Transfection</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>eNqFkcFvFCEYxSdGU9fq2Ysa4kEvnS0fMAwkXky1alLjQXsmDMOsrCxMgTHtf-9sdrtaD3oiX97vPeB7VfUU8BJwS0_HoPNSwhJgyVoG96oFYAk1ZxLfrxYYk7YWjLCH1aOc1xhj2Qh8VB0JkC0Qsajy5-itmbxOyPgYXFidoOJynizqXS7JdVNxMZwgHXpkr8dkc55nFAekEbD6xzuNOuct0sb1dedCP0egMcViXUBDihuUdEEzoT0yNyXm6B9XDwbts32yP4-ry_P3384-1hdfPnw6e3tRG45lqQVvcEcYJ1bLhjHGOys5Y4YOorEcW2paanUPQ2MpkdjiroVWaNKRpjOmIfS4erPLHaduY3tjQ0naqzG5jU43Kmqn7irBfVer-FMxKjib7a_29hSvJpuL2rhsrPc62Dhl1fKGEC7Jf0HgQgATcgZf_gWu45TCvANFMFDaMkpn6HQHmRRzTnY4PBiw2nautp0rCQpAbTufHc___OeB35c86y_2-tZ4q94JeP1PQA2T98Vel5l8tiPXucR0QAnlTLDm902Djkqvksvq8itI2WBMMW0x_QX1_dMh</recordid><startdate>19940524</startdate><enddate>19940524</enddate><creator>Gong, Yong-Zhong</creator><creator>Everett, Eric T.</creator><creator>Schwartz, David A.</creator><creator>Norris, James S.</creator><creator>Wilson, Frederick A.</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>FBQ</scope><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>19940524</creationdate><title>Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol</title><author>Gong, Yong-Zhong ; Everett, Eric T. ; Schwartz, David A. ; Norris, James S. ; Wilson, Frederick A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c609t-8650b2462ea954446be9644c3f85e60e3c73ead1f5e3290e0b7178a2b25bcc523</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>acide biliaire</topic><topic>acidos biliares</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Animals</topic><topic>arn mensajero</topic><topic>arn messager</topic><topic>Base Sequence</topic><topic>Bile</topic><topic>Bile acids</topic><topic>Bile Acids and Salts - metabolism</topic><topic>binding proteins</topic><topic>Biochemistry</topic><topic>Blotting, Northern</topic><topic>Blotting, Western</topic><topic>Carrier Proteins - biosynthesis</topic><topic>Carrier Proteins - genetics</topic><topic>Carrier Proteins - metabolism</topic><topic>Cell Line</topic><topic>Cells</topic><topic>citoplasma</topic><topic>Cloning, Molecular</topic><topic>Complementary DNA</topic><topic>COS cells</topic><topic>cytoplasm</topic><topic>cytoplasme</topic><topic>Cytosol - metabolism</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>expresion genica</topic><topic>expression des genes</topic><topic>gene</topic><topic>gene expression</topic><topic>genes</topic><topic>Gongs</topic><topic>Humans</topic><topic>Hydroxysteroid Dehydrogenases</topic><topic>Ileum</topic><topic>Ileum - metabolism</topic><topic>intestin</topic><topic>intestines</topic><topic>intestinos</topic><topic>Liver</topic><topic>Membrane Glycoproteins</topic><topic>messenger rna</topic><topic>Molecular Sequence Data</topic><topic>nucleotide sequence</topic><topic>Organ Specificity - genetics</topic><topic>proteinas aglutinantes</topic><topic>proteine de liaison</topic><topic>Proteins</topic><topic>rat</topic><topic>rata</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>RNA</topic><topic>Rodents</topic><topic>secuencia nucleica</topic><topic>Sequence Homology, Amino Acid</topic><topic>sequence nucleique</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gong, Yong-Zhong</creatorcontrib><creatorcontrib>Everett, Eric T.</creatorcontrib><creatorcontrib>Schwartz, David A.</creatorcontrib><creatorcontrib>Norris, James S.</creatorcontrib><creatorcontrib>Wilson, Frederick A.</creatorcontrib><collection>AGRIS</collection><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>Gong, Yong-Zhong</au><au>Everett, Eric T.</au><au>Schwartz, David A.</au><au>Norris, James S.</au><au>Wilson, Frederick A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1994-05-24</date><risdate>1994</risdate><volume>91</volume><issue>11</issue><spage>4741</spage><epage>4745</epage><pages>4741-4745</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>A cDNA clone encoding the major intestinal cytosolic 14-kDa bile acid-binding protein (14-kDa I-BABP) was isolated from a rat ileal lambda gt22A library following immunoscreening using a monospecific antiserum raised against a 14-kDa polypeptide found in the rat ileal cytosol. One done of 516 bp encoded a 128-amino acid protein with a predicted molecular mass of 14,544 Da. The deduced amino acid sequence of 14-kDa I-BABP showed 100% homology to rat intestinal 15-kDa protein (I-15P) and 72% homology to porcine 15-kDa gastrotropin, whereas comparison of I-BABP to rat 14-kDa fatty acid-binding proteins of liver, intestine, and heart revealed homologies of 44%, 25%, and 28%, respectively. Northern blot analysis revealed a single transcript of approximately 0.5 kb in ileum and ovary; however, the abundance of I-BABP mRNA was much greater in ileum than in ovary. No transcript was seen in RNA extracted from stomach. jejunum, colon, liver, adrenal, brain, heart, kidney, or testis. Transfection of the I-BABP cDNA into COS-7 cells resulted in the expression of a 14-kDa protein that was identical to the ileal cytosolic I-BABP as determined by immunoblotting. Photoaffinity labeling of expressed 14-kDa protein was saturable with respect to increasing concentrations of 7,7-azo[(3)H]taurocholate [K(m), 83.3 micromolars; V(max), 6.7 pmol/mg per 5 min]. Taurocholate inhibited 7,7-azotaurocholate labeling by >96% with lesser inhibition by taurochenodeoxycholate (83.1%), chenodeoxycholate (74.6%), cholate (50.5%), and progesterone (38.5%), whereas oleic acid and estradiol did not inhibit binding.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>8197128</pmid><doi>10.1073/pnas.91.11.4741</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1994-05, Vol.91 (11), p.4741-4745 |
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| subjects | acide biliaire acidos biliares Amino Acid Sequence Amino acids Animals arn mensajero arn messager Base Sequence Bile Bile acids Bile Acids and Salts - metabolism binding proteins Biochemistry Blotting, Northern Blotting, Western Carrier Proteins - biosynthesis Carrier Proteins - genetics Carrier Proteins - metabolism Cell Line Cells citoplasma Cloning, Molecular Complementary DNA COS cells cytoplasm cytoplasme Cytosol - metabolism Deoxyribonucleic acid DNA expresion genica expression des genes gene gene expression genes Gongs Humans Hydroxysteroid Dehydrogenases Ileum Ileum - metabolism intestin intestines intestinos Liver Membrane Glycoproteins messenger rna Molecular Sequence Data nucleotide sequence Organ Specificity - genetics proteinas aglutinantes proteine de liaison Proteins rat rata Rats Rats, Sprague-Dawley RNA Rodents secuencia nucleica Sequence Homology, Amino Acid sequence nucleique Transfection |
| title | Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol |
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