New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA
Identifieur interne : 001C45 ( Istex/Corpus ); précédent : 001C44; suivant : 001C46New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA
Auteurs : Marc Desrochers ; Maro Christou ; Colin Jefcoate ; Anne Belzil ; Alan AndersonSource :
- Biochemical Pharmacology [ 0006-2952 ] ; 1996.
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- KwdEn :
Abstract
The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[a]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.
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DOI: 10.1016/0006-2952(96)00502-3
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title><author><name sortKey="Desrochers, Marc" sort="Desrochers, Marc" uniqKey="Desrochers M" first="Marc" last="Desrochers">Marc Desrochers</name><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation><affiliation><mods:affiliation>‡Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</mods:affiliation></affiliation></author><author><name sortKey="Christou, Maro" sort="Christou, Maro" uniqKey="Christou M" first="Maro" last="Christou">Maro Christou</name><affiliation><mods:affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</mods:affiliation></affiliation></author><author><name sortKey="Jefcoate, Colin" sort="Jefcoate, Colin" uniqKey="Jefcoate C" first="Colin" last="Jefcoate">Colin Jefcoate</name><affiliation><mods:affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</mods:affiliation></affiliation></author><author><name sortKey="Belzil, Anne" sort="Belzil, Anne" uniqKey="Belzil A" first="Anne" last="Belzil">Anne Belzil</name><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation></author><author><name sortKey="Anderson, Alan" sort="Anderson, Alan" uniqKey="Anderson A" first="Alan" last="Anderson">Alan Anderson</name><affiliation><mods:affiliation>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:2CACE07AB3E8374A86F3F446CAB80F572849CAFE</idno><date when="1996" year="1996">1996</date><idno type="doi">10.1016/0006-2952(96)00502-3</idno><idno type="url">https://api.istex.fr/document/2CACE07AB3E8374A86F3F446CAB80F572849CAFE/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">001C45</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001C45</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title><author><name sortKey="Desrochers, Marc" sort="Desrochers, Marc" uniqKey="Desrochers M" first="Marc" last="Desrochers">Marc Desrochers</name><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation><affiliation><mods:affiliation>‡Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</mods:affiliation></affiliation></author><author><name sortKey="Christou, Maro" sort="Christou, Maro" uniqKey="Christou M" first="Maro" last="Christou">Maro Christou</name><affiliation><mods:affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</mods:affiliation></affiliation></author><author><name sortKey="Jefcoate, Colin" sort="Jefcoate, Colin" uniqKey="Jefcoate C" first="Colin" last="Jefcoate">Colin Jefcoate</name><affiliation><mods:affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</mods:affiliation></affiliation></author><author><name sortKey="Belzil, Anne" sort="Belzil, Anne" uniqKey="Belzil A" first="Anne" last="Belzil">Anne Belzil</name><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation></author><author><name sortKey="Anderson, Alan" sort="Anderson, Alan" uniqKey="Anderson A" first="Alan" last="Anderson">Alan Anderson</name><affiliation><mods:affiliation>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</mods:affiliation></affiliation><affiliation><mods:affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Biochemical Pharmacology</title><title level="j" type="abbrev">BCP</title><idno type="ISSN">0006-2952</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1996">1996</date><biblScope unit="volume">52</biblScope><biblScope unit="issue">8</biblScope><biblScope unit="page" from="1311">1311</biblScope><biblScope unit="page" to="1319">1319</biblScope></imprint><idno type="ISSN">0006-2952</idno></series><idno type="istex">2CACE07AB3E8374A86F3F446CAB80F572849CAFE</idno><idno type="DOI">10.1016/0006-2952(96)00502-3</idno><idno type="PII">0006-2952(96)00502-3</idno><idno type="ArticleID">96005023</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-2952</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>CYP2B</term><term>alternative splicing</term><term>antipeptide antibodies</term><term>cytochrome P450</term><term>phenobarbital</term><term>rat liver</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[a]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Marc Desrochers</name><affiliations><json:string>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</json:string><json:string>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</json:string><json:string>‡Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</json:string></affiliations></json:item><json:item><name>Maro Christou</name><affiliations><json:string>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</json:string></affiliations></json:item><json:item><name>Colin Jefcoate</name><affiliations><json:string>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</json:string></affiliations></json:item><json:item><name>Anne Belzil</name><affiliations><json:string>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</json:string><json:string>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</json:string></affiliations></json:item><json:item><name>Alan Anderson</name><affiliations><json:string>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</json:string><json:string>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</json:string><json:string>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>phenobarbital</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>CYP2B</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>cytochrome P450</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>alternative splicing</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>antipeptide antibodies</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>rat liver</value></json:item></subject><articleId><json:string>96005023</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[a]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.</abstract><qualityIndicators><score>7.952</score><pdfVersion>1.2</pdfVersion><pdfPageSize>576 x 792 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>6</keywordCount><abstractCharCount>1760</abstractCharCount><pdfWordCount>5630</pdfWordCount><pdfCharCount>38806</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>246</abstractWordCount></qualityIndicators><title>New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title><pii><json:string>0006-2952(96)00502-3</json:string></pii><refBibs><json:item><author><json:item><name>FJ Gonzalez</name></json:item></author><host><volume>40</volume><pages><last>288</last><first>243</first></pages><author></author><title>Pharmocol Rev</title></host><title>The molecular biology of cytochrome P450s</title></json:item><json:item><author><json:item><name>TD Porter</name></json:item><json:item><name>MJ Coon</name></json:item></author><host><volume>266</volume><pages><last>13472</last><first>13469</first></pages><author></author><title>J Biol Chem</title></host><title>Cytochrome P-450. 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pharmacy</json:string></scienceMetrix></categories><publicationDate>1996</publicationDate><copyrightDate>1996</copyrightDate><doi><json:string>10.1016/0006-2952(96)00502-3</json:string></doi><id>2CACE07AB3E8374A86F3F446CAB80F572849CAFE</id><score>0.07014032</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/document/2CACE07AB3E8374A86F3F446CAB80F572849CAFE/fulltext/pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/document/2CACE07AB3E8374A86F3F446CAB80F572849CAFE/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/2CACE07AB3E8374A86F3F446CAB80F572849CAFE/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>ELSEVIER</publisher><availability><p>ELSEVIER</p></availability><date>1996</date></publicationStmt><notesStmt><note>This work was supported by grants from the Medical Research Council of Canada (MA-8733 to A.A.) and from the U.S. National Institutes of Health (CA16265 toC.J.).</note><note type="content">Section title: Research paper</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title><author xml:id="author-1"><persName><forename type="first">Marc</forename><surname>Desrochers</surname></persName><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation><affiliation>‡Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</affiliation></author><author xml:id="author-2"><persName><forename type="first">Maro</forename><surname>Christou</surname></persName><affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</affiliation></author><author xml:id="author-3"><persName><forename type="first">Colin</forename><surname>Jefcoate</surname></persName><affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</affiliation></author><author xml:id="author-4"><persName><forename type="first">Anne</forename><surname>Belzil</surname></persName><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation></author><author xml:id="author-5"><persName><forename type="first">Alan</forename><surname>Anderson</surname></persName><affiliation>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</affiliation><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation></author></analytic><monogr><title level="j">Biochemical Pharmacology</title><title level="j" type="abbrev">BCP</title><idno type="pISSN">0006-2952</idno><idno type="PII">S0006-2952(00)X0041-X</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1996"></date><biblScope unit="volume">52</biblScope><biblScope unit="issue">8</biblScope><biblScope unit="page" from="1311">1311</biblScope><biblScope unit="page" to="1319">1319</biblScope></imprint></monogr><idno type="istex">2CACE07AB3E8374A86F3F446CAB80F572849CAFE</idno><idno type="DOI">10.1016/0006-2952(96)00502-3</idno><idno type="PII">0006-2952(96)00502-3</idno><idno type="ArticleID">96005023</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1996</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[a]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>Keywords</head><item><term>phenobarbital</term></item><item><term>CYP2B</term></item><item><term>cytochrome P450</term></item><item><term>alternative splicing</term></item><item><term>antipeptide antibodies</term></item><item><term>rat liver</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1996">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/2CACE07AB3E8374A86F3F446CAB80F572849CAFE/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>BCP</jid><aid>96005023</aid><ce:pii>0006-2952(96)00502-3</ce:pii><ce:doi>10.1016/0006-2952(96)00502-3</ce:doi><ce:copyright type="unknown" year="1996"></ce:copyright></item-info><head><ce:article-footnote><ce:label>☆</ce:label><ce:note-para>This work was supported by grants from the Medical Research Council of Canada (MA-8733 to A.A.) and from the U.S. National Institutes of Health (CA16265 toC.J.).</ce:note-para></ce:article-footnote><ce:dochead><ce:textfn>Research paper</ce:textfn></ce:dochead><ce:title>New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</ce:title><ce:author-group><ce:author><ce:given-name>Marc</ce:given-name><ce:surname>Desrochers</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup loc="post">†</ce:sup></ce:cross-ref><ce:cross-ref refid="FN1"><ce:sup loc="post">‡</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Maro</ce:given-name><ce:surname>Christou</ce:surname><ce:cross-ref refid="AFF3"><ce:sup loc="post">§</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Colin</ce:given-name><ce:surname>Jefcoate</ce:surname><ce:cross-ref refid="AFF3"><ce:sup loc="post">§</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Anne</ce:given-name><ce:surname>Belzil</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup loc="post">†</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Alan</ce:given-name><ce:surname>Anderson</ce:surname><ce:cross-ref refid="COR1"><ce:sup loc="post">∥</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup loc="post">†</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>∥</ce:label><ce:text>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</ce:text></ce:correspondence><ce:footnote id="FN1"><ce:label>‡</ce:label><ce:note-para>Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="29" month="12" year="1995"></ce:date-received><ce:date-accepted day="20" month="5" year="1996"></ce:date-accepted><ce:abstract class="author"><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para view="all" id="simple-para.0010">The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[<ce:italic>a</ce:italic>]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>phenobarbital</ce:text></ce:keyword><ce:keyword><ce:text>CYP2B</ce:text></ce:keyword><ce:keyword><ce:text>cytochrome P450</ce:text></ce:keyword><ce:keyword><ce:text>alternative splicing</ce:text></ce:keyword><ce:keyword><ce:text>antipeptide antibodies</ce:text></ce:keyword><ce:keyword><ce:text>rat liver</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>New proteins in the rat CYP2B subfamily: Presence in liver microsomes of the constitutive CYP2B3 protein and the phenobarbital-inducible protein product of alternatively spliced CYP2B2 mRNA</title></titleInfo><name type="personal"><namePart type="given">Marc</namePart><namePart type="family">Desrochers</namePart><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation><affiliation>‡Present address: INSERM U347, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre, France.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Maro</namePart><namePart type="family">Christou</namePart><affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Colin</namePart><namePart type="family">Jefcoate</namePart><affiliation>Department of Pharmacology, University of Wisconsin Medical School, Madison, WI 53706, U.S.A.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Anne</namePart><namePart type="family">Belzil</namePart><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Alan</namePart><namePart type="family">Anderson</namePart><affiliation>Corresponding author: Alan Anderson, Ph.D., Centre de recherche, L'Hôtel-Dieu de Québec, 11 côte du Palais, Québec, Canada G1R2J6. Tel. (418) 691-5548; FAX (418) 691-5439.</affiliation><affiliation>Centre De Recherche En CancÉrologie De L' UniversitÉ Lav Al, L' HÔtel-Dieu De QuÉbec, Centre Hospitalier Universitaire De Québec, QUébec, Canada G1R2J6</affiliation><affiliation>Départment De Biologie, Université Laval, Québec, Canada G1K7P4</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1996</dateIssued><copyrightDate encoding="w3cdtf">1996</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">The rat CYP2B gene subfamily includes CYP2B1, CYP2B2 and CYP2B3. Translation of an alternatively spliced hepatic CYP2B2 mRNA would generate a CYP2B2 variant, CYP2B2v, having eight additional amino acid residues inserted between CYP2B2 positions 274 and 275. The presence of CYP2B3 and CYP2B2v in rat liver has yet to be demonstrated. cDNA expression vectors were obtained for CYP2B1, CYP2B2, CYP2B3 and CYP2B2v. All four proteins react with an anti-CYP2Bl antibody and can be resolved by SDS-PAGE. A CYP2B3-specific polyclonal antibody raised against an undecapeptide (SPVDPNTIDMT) from near the C-terminus of CYP2B3 detected a constitutive protein on immunoblots of rat liver microsomes, thus demonstrating that the CYP2B3 mRNA is translated in the liver. Similarly, a CYP2B2v-specific polyclonal antibody was raised against a peptide containing the eight additional amino acid residues (VSPAWMRE) predicted to be present in the CYP2B2v protein. It detected a phenobarbital- and Aroclor 1254-inducible protein in rat liver microsomes. Microsomes of Ad293 cells expressing cDNAs for CYP2B2 and CYP2B2v were used to metabolize 7,12-dimethylbenz[a]anthracene (DMBA), and the metabolites produced were compared with those generated by microsomes of cells expressing CYP2B1 cDNA. CYP2B2v had activity similar to that of CYP2B2 for DMBA metabolism. Both CYP2B2 forms preferentially catalyzed 12-hydroxylation, whereas CYP2B1 preferred 7-hydroxylation and exhibited turnover that was strongly suppressed as previously reported. These results demonstrate the existence in rat liver of two new CYP2B proteins: CYP2B3, the major constitutive CYP2B form, and CYP2B2v, which represents a rare case of non-aberrant alternative splicing among xenobiotic-metabolizing P450s.</abstract><note>This work was supported by grants from the Medical Research Council of Canada (MA-8733 to A.A.) and from the U.S. National Institutes of Health (CA16265 toC.J.).</note><note type="content">Section title: Research paper</note><subject lang="en"><genre>Keywords</genre><topic>phenobarbital</topic><topic>CYP2B</topic><topic>cytochrome P450</topic><topic>alternative splicing</topic><topic>antipeptide antibodies</topic><topic>rat liver</topic></subject><relatedItem type="host"><titleInfo><title>Biochemical Pharmacology</title></titleInfo><titleInfo type="abbreviated"><title>BCP</title></titleInfo><genre type="journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">19961025</dateIssued></originInfo><identifier type="ISSN">0006-2952</identifier><identifier type="PII">S0006-2952(00)X0041-X</identifier><part><date>19961025</date><detail type="volume"><number>52</number><caption>vol.</caption></detail><detail type="issue"><number>8</number><caption>no.</caption></detail><extent unit="issue pages"><start>1133</start><end>1321</end></extent><extent unit="pages"><start>1311</start><end>1319</end></extent></part></relatedItem><identifier type="istex">2CACE07AB3E8374A86F3F446CAB80F572849CAFE</identifier><identifier type="DOI">10.1016/0006-2952(96)00502-3</identifier><identifier type="PII">0006-2952(96)00502-3</identifier><identifier type="ArticleID">96005023</identifier><recordInfo><recordContentSource>ELSEVIER</recordContentSource></recordInfo></mods></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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