RGD Reference Report - Purification and characterization of recombinant rat hepatic CYP4F1. - Rat Genome Database

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Purification and characterization of recombinant rat hepatic CYP4F1.

Authors: Kikuta, Y  Kusunose, E  Ito, M  Kusunose, M 
Citation: Kikuta Y, etal., Arch Biochem Biophys. 1999 Sep 15;369(2):193-6.
RGD ID: 2301713
Pubmed: PMID:10486137   (View Abstract at PubMed)
DOI: DOI:10.1006/abbi.1999.1271   (Journal Full-text)

CYP4F1 was discovered by Chen and Hardwick (Arch. Biochem. Biophys. 300, 18-23, 1993) as a new CYP4 cytochrome P450 (P450) preferentially expressed in rat hepatomas. However, the catalytic function of this P450 remained poorly defined. We have purified recombinant CYP4F1 protein to a specific content of 12 nmol of P450/mg of protein from transfected yeast cells by chromatography of solubilized microsomes on an amino-n-hexyl Sepharose 4B column, followed by sequential HPLC on a DEAE column and two hydroxylapatite columns. The purified P450 was homogeneous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an apparent molecular weight of 53 kDa. The enzyme catalyzed the omega-hydroxylation of leukotriene B(4) with a K(m) of 134 microM and a V(max) of 6.5 nmol/min/nmol of P450 in the presence of rabbit hepatic NADPH-P450 reductase and cytochrome b(5). In addition, 6-trans-LTB(4), lipoxin A(4), prostaglandin A(1), and several hydroxyeicosatetraenoic acids (HETEs) were also omega-hydroxylated. Of several eicosanoids examined, 8-HETE was the most efficient substrate, with a K(m) of 18.6 microM and a V(max) of 15.8 nmol/min/nmol of P450. In contrast, no activity was detected toward lipoxin B(4), laurate, palmitate, arachidonate, and benzphetamine. The results suggest that CYP4F1 participates in the hepatic inactivation of several bioactive eicosanoids.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Cyp4f1Raticosanoid metabolic process  IDA  RGD 
Cyp4f1Ratleukotriene B4 catabolic process involved_inIDA PMID:10486137UniProt 

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Cyp4f1Ratleukotriene-B4 20-monooxygenase activity enablesIDA PMID:10486137UniProt 

Objects Annotated

Genes (Rattus norvegicus)
Cyp4f1  (cytochrome P450, family 4, subfamily f, polypeptide 1)


Additional Information