Molecular cloning, expression, and enzymatic characterization of the rat kidney cytochrome P-450 arachidonic acid epoxygenase
A cDNA containing an open reading frame coding for the rat kidney cytochrome P-450 arachidonic acid epoxygenase was isolated from a male rat kidney cDNA library. Sequence analysis showed that with the exception of 11 nucleotides, this cDNA is identical with the published sequence for rat liver cytoc...
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| Veröffentlicht in: | The Journal of biological chemistry 1993-06, Vol.268 (18), p.13565-13570 |
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| container_title | The Journal of biological chemistry |
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| creator | KARARA, A MAKITA, K JACOBSON, H. R FALCK, J. R GUENGERICH, F. P DUBOIS, R. N CAPDEVILA, J. H |
| description | A cDNA containing an open reading frame coding for the rat kidney cytochrome P-450 arachidonic acid epoxygenase was isolated
from a male rat kidney cDNA library. Sequence analysis showed that with the exception of 11 nucleotides, this cDNA is identical
with the published sequence for rat liver cytochrome 2C23 and encodes a polypeptide of 494 amino acids. Nucleic acid blot
hybridization indicated that the levels of expression of the corresponding mRNA are high in rat kidney and liver and are undetectable
in brain and heart. The cDNA coding region was cloned into a pCMV2 vector and expressed in COS-1 cells. The recombinant microsomal
protein catalyzed the NADPH-dependent metabolism of arachidonic acid to a mixture of 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic
acids as the only oxygenation products. The enantiofacial selectivity of the recombinant protein was nearly identical with
that reported for the kidney microsomal enzyme and generated 8(R),9(S)-, 11(R),12(S)-, and 14(S),15(R) with optical purities
of 95, 85, and 75%, respectively. On the basis of mRNA abundance and the close similarities between the regio- and stereochemical
selectivity of the recombinant and kidney microsomal proteins, we concluded that cytochrome P-450 2C23 is the predominant
enzyme isoform responsible for arachidonic acid epoxidation in the rat kidney. |
| doi_str_mv | 10.1016/s0021-9258(19)38686-7 |
| format | Article |
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from a male rat kidney cDNA library. Sequence analysis showed that with the exception of 11 nucleotides, this cDNA is identical
with the published sequence for rat liver cytochrome 2C23 and encodes a polypeptide of 494 amino acids. Nucleic acid blot
hybridization indicated that the levels of expression of the corresponding mRNA are high in rat kidney and liver and are undetectable
in brain and heart. The cDNA coding region was cloned into a pCMV2 vector and expressed in COS-1 cells. The recombinant microsomal
protein catalyzed the NADPH-dependent metabolism of arachidonic acid to a mixture of 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic
acids as the only oxygenation products. The enantiofacial selectivity of the recombinant protein was nearly identical with
that reported for the kidney microsomal enzyme and generated 8(R),9(S)-, 11(R),12(S)-, and 14(S),15(R) with optical purities
of 95, 85, and 75%, respectively. On the basis of mRNA abundance and the close similarities between the regio- and stereochemical
selectivity of the recombinant and kidney microsomal proteins, we concluded that cytochrome P-450 2C23 is the predominant
enzyme isoform responsible for arachidonic acid epoxidation in the rat kidney.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/s0021-9258(19)38686-7</identifier><identifier>PMID: 8514789</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Animals ; Biological and medical sciences ; Cells, Cultured ; Cloning, Molecular ; Cytochrome P-450 Enzyme System - genetics ; Cytochrome P-450 Enzyme System - metabolism ; DNA - isolation & purification ; Eicosanoids - metabolism ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Kidney - enzymology ; Male ; Microsomes - metabolism ; Molecular Sequence Data ; NADP - metabolism ; Oxidoreductases ; Oxygenases - genetics ; Oxygenases - metabolism ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism</subject><ispartof>The Journal of biological chemistry, 1993-06, Vol.268 (18), p.13565-13570</ispartof><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c506t-af123e8773bfec73bc36feabc11d89d65bc0e4cd5dd08868cad19b831e4aecb53</citedby><cites>FETCH-LOGICAL-c506t-af123e8773bfec73bc36feabc11d89d65bc0e4cd5dd08868cad19b831e4aecb53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4829753$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8514789$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KARARA, A</creatorcontrib><creatorcontrib>MAKITA, K</creatorcontrib><creatorcontrib>JACOBSON, H. R</creatorcontrib><creatorcontrib>FALCK, J. R</creatorcontrib><creatorcontrib>GUENGERICH, F. P</creatorcontrib><creatorcontrib>DUBOIS, R. N</creatorcontrib><creatorcontrib>CAPDEVILA, J. H</creatorcontrib><title>Molecular cloning, expression, and enzymatic characterization of the rat kidney cytochrome P-450 arachidonic acid epoxygenase</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>A cDNA containing an open reading frame coding for the rat kidney cytochrome P-450 arachidonic acid epoxygenase was isolated
from a male rat kidney cDNA library. Sequence analysis showed that with the exception of 11 nucleotides, this cDNA is identical
with the published sequence for rat liver cytochrome 2C23 and encodes a polypeptide of 494 amino acids. Nucleic acid blot
hybridization indicated that the levels of expression of the corresponding mRNA are high in rat kidney and liver and are undetectable
in brain and heart. The cDNA coding region was cloned into a pCMV2 vector and expressed in COS-1 cells. The recombinant microsomal
protein catalyzed the NADPH-dependent metabolism of arachidonic acid to a mixture of 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic
acids as the only oxygenation products. The enantiofacial selectivity of the recombinant protein was nearly identical with
that reported for the kidney microsomal enzyme and generated 8(R),9(S)-, 11(R),12(S)-, and 14(S),15(R) with optical purities
of 95, 85, and 75%, respectively. On the basis of mRNA abundance and the close similarities between the regio- and stereochemical
selectivity of the recombinant and kidney microsomal proteins, we concluded that cytochrome P-450 2C23 is the predominant
enzyme isoform responsible for arachidonic acid epoxidation in the rat kidney.</description><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Cloning, Molecular</subject><subject>Cytochrome P-450 Enzyme System - genetics</subject><subject>Cytochrome P-450 Enzyme System - metabolism</subject><subject>DNA - isolation & purification</subject><subject>Eicosanoids - metabolism</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Kidney - enzymology</subject><subject>Male</subject><subject>Microsomes - metabolism</subject><subject>Molecular Sequence Data</subject><subject>NADP - metabolism</subject><subject>Oxidoreductases</subject><subject>Oxygenases - genetics</subject><subject>Oxygenases - metabolism</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkVFrFDEUhYModa3-hEIeRCx0NJlMZpJHKVaFioIKvoXMzZ2d6EyyTWaxW_C_m7XL-mgIuYT7nXPhXELOOHvFGW9fZ8ZqXulaqpdcnwvVqrbqHpAVZ0pUQvLvD8nqiDwmT3L-wcppND8hJ0ryplN6RX5_jBPCdrKJwhSDD-sLirebhDn7GC6oDY5iuNvNdvFAYbTJwoLJ35V_DDQOdBmRJrvQn94F3FHYLRHGFGekn6tGMrpXjN4Vb6AWfLHbxNvdGoPN-JQ8GuyU8dmhnpJvV2-_Xr6vrj-9-3D55roCydqlsgOvBaquE_2AUF4Q7YC2B86d0q6VPTBswEnnmCpBgHVc90pwbCxCL8UpeXHvu0nxZot5MbPPgNNkA8ZtNp3stGjq5r8gb1utmWYFlPcgpJhzwsFskp9t2hnOzH4_5ss-fLMP33Bt_u7HdEV3dhiw7Wd0R9VhIaX__NC3Gew0JBvA5yPWqFp3UvzDRr8ef_mEpvcldpxN3ZZ55QrZSvEHCY-nJw</recordid><startdate>19930625</startdate><enddate>19930625</enddate><creator>KARARA, A</creator><creator>MAKITA, K</creator><creator>JACOBSON, H. R</creator><creator>FALCK, J. R</creator><creator>GUENGERICH, F. P</creator><creator>DUBOIS, R. N</creator><creator>CAPDEVILA, J. H</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>IQODW</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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19930625</creationdate><title>Molecular cloning, expression, and enzymatic characterization of the rat kidney cytochrome P-450 arachidonic acid epoxygenase</title><author>KARARA, A ; MAKITA, K ; JACOBSON, H. R ; FALCK, J. R ; GUENGERICH, F. P ; DUBOIS, R. N ; CAPDEVILA, J. H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c506t-af123e8773bfec73bc36feabc11d89d65bc0e4cd5dd08868cad19b831e4aecb53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>Cloning, Molecular</topic><topic>Cytochrome P-450 Enzyme System - genetics</topic><topic>Cytochrome P-450 Enzyme System - metabolism</topic><topic>DNA - isolation & purification</topic><topic>Eicosanoids - metabolism</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Kidney - enzymology</topic><topic>Male</topic><topic>Microsomes - metabolism</topic><topic>Molecular Sequence Data</topic><topic>NADP - metabolism</topic><topic>Oxidoreductases</topic><topic>Oxygenases - genetics</topic><topic>Oxygenases - metabolism</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KARARA, A</creatorcontrib><creatorcontrib>MAKITA, K</creatorcontrib><creatorcontrib>JACOBSON, H. R</creatorcontrib><creatorcontrib>FALCK, J. R</creatorcontrib><creatorcontrib>GUENGERICH, F. P</creatorcontrib><creatorcontrib>DUBOIS, R. N</creatorcontrib><creatorcontrib>CAPDEVILA, J. H</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KARARA, A</au><au>MAKITA, K</au><au>JACOBSON, H. R</au><au>FALCK, J. R</au><au>GUENGERICH, F. P</au><au>DUBOIS, R. N</au><au>CAPDEVILA, J. H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning, expression, and enzymatic characterization of the rat kidney cytochrome P-450 arachidonic acid epoxygenase</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1993-06-25</date><risdate>1993</risdate><volume>268</volume><issue>18</issue><spage>13565</spage><epage>13570</epage><pages>13565-13570</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>A cDNA containing an open reading frame coding for the rat kidney cytochrome P-450 arachidonic acid epoxygenase was isolated
from a male rat kidney cDNA library. Sequence analysis showed that with the exception of 11 nucleotides, this cDNA is identical
with the published sequence for rat liver cytochrome 2C23 and encodes a polypeptide of 494 amino acids. Nucleic acid blot
hybridization indicated that the levels of expression of the corresponding mRNA are high in rat kidney and liver and are undetectable
in brain and heart. The cDNA coding region was cloned into a pCMV2 vector and expressed in COS-1 cells. The recombinant microsomal
protein catalyzed the NADPH-dependent metabolism of arachidonic acid to a mixture of 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic
acids as the only oxygenation products. The enantiofacial selectivity of the recombinant protein was nearly identical with
that reported for the kidney microsomal enzyme and generated 8(R),9(S)-, 11(R),12(S)-, and 14(S),15(R) with optical purities
of 95, 85, and 75%, respectively. On the basis of mRNA abundance and the close similarities between the regio- and stereochemical
selectivity of the recombinant and kidney microsomal proteins, we concluded that cytochrome P-450 2C23 is the predominant
enzyme isoform responsible for arachidonic acid epoxidation in the rat kidney.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>8514789</pmid><doi>10.1016/s0021-9258(19)38686-7</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1993-06, Vol.268 (18), p.13565-13570 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_75793424 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Cells, Cultured Cloning, Molecular Cytochrome P-450 Enzyme System - genetics Cytochrome P-450 Enzyme System - metabolism DNA - isolation & purification Eicosanoids - metabolism Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Gene Expression Kidney - enzymology Male Microsomes - metabolism Molecular Sequence Data NADP - metabolism Oxidoreductases Oxygenases - genetics Oxygenases - metabolism Rats Rats, Sprague-Dawley Recombinant Proteins - genetics Recombinant Proteins - metabolism |
| title | Molecular cloning, expression, and enzymatic characterization of the rat kidney cytochrome P-450 arachidonic acid epoxygenase |
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