| Structural determination of the substrate specificities and regioselectivities of the rat and human fatty acid omega-hydroxylases. | |
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MedLine Citation:
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PMID: 10620324 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The substrate and regiospecificities of the known CYP4A enzymes from rat (CYP4A1, -4A2, -4A3, and -4A8) and human (CYP4A11) have been determined using lauric (C12), myristic (C14), palmitic (C16), oleic (C18:1), and arachidonic (C20:4) acids. The CYP4A2 and CYP4A8 cDNAs required to complete the enzyme set were cloned from a rat kidney library. All five proteins were expressed in Escherichia coli and were purified with the help of a six-histidine tag at the carboxyl terminus. Two complementary CYP4A2-CYP4A3 chimeras fused at residue 119 (CYP4A2) and 122 (CYP4A3) were constructed to explore the roles of the 18 amino acid differences between the parent proteins in determining their catalytic profiles. The chimera in which the first 119 amino acids are from CYP4A2 indicates that the first 120 amino acids control the substrate specificity. The chimera in which the first 122 amino acids are from CYP4A3 is inactive due to a defect in electron transfer to the heme group. The highest activity for lauric acid was obtained with CYP4A1 and CYP4A8, but for all the proteins the activity decreased with increasing fatty acid chain length. The fact that none of the rat and human CYP4A enzymes exhibits a high activity with arachidonic acid appears to limit their role as catalysts for the physiologically important conversion of arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE). |
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Authors:
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U Hoch; Z Zhang; D L Kroetz; P R Ortiz de Montellano |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Archives of biochemistry and biophysics Volume: 373 ISSN: 0003-9861 ISO Abbreviation: Arch. Biochem. Biophys. Publication Date: 2000 Jan |
Date Detail:
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Created Date: 2000-02-09 Completed Date: 2000-02-09 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0372430 Medline TA: Arch Biochem Biophys Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 63-71 Citation Subset: IM |
Copyright Information:
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Copyright 2000 Academic Press. |
Affiliation:
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Department of Pharmaceutical Chemistry, University of California, San Francisco, California, 94143-0446, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Alkane 1-Monooxygenase Amino Acid Sequence Animals Base Sequence Catalytic Domain / genetics Cytochrome P-450 Enzyme System / chemistry*, genetics, metabolism* DNA Primers / genetics Escherichia coli / genetics Fatty Acids Humans Isoenzymes / chemistry, genetics, metabolism Mixed Function Oxygenases / chemistry*, genetics, metabolism* Models, Molecular Molecular Sequence Data Protein Conformation Rats Recombinant Fusion Proteins / chemistry, genetics, metabolism Recombinant Proteins / chemistry, genetics, metabolism Sequence Homology, Amino Acid Spectrophotometry Substrate Specificity |
| Grant Support | |
ID/Acronym/Agency:
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GM25515/GM/NIGMS NIH HHS; HL53994/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/Fatty Acids; 0/Isoenzymes; 0/Recombinant Fusion Proteins; 0/Recombinant Proteins; 9035-51-2/Cytochrome P-450 Enzyme System; EC 1.-/Mixed Function Oxygenases; EC 1.14.15.3/Alkane 1-Monooxygenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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