Expression and Catalytic Activity of Mouse Leukotriene B4 ω-Hydroxylase, CYP4F14

Autor: Yasushi Kikuta, Masamichi Kusunose, Emi Kusunose, Hideaki Kasyu
Rok vydání: 2000
Předmět:
Male
Transcription
Genetic
Leukotriene B4
Biochemistry
Mixed Function Oxygenases
Substrate Specificity
Mice
chemistry.chemical_compound
Exon
Cytochrome P-450 Enzyme System
Hydroxyeicosatetraenoic Acids
Tissue Distribution
Clofibrate
Mice
Knockout
Arachidonic Acid
Expression vector
Reverse Transcriptase Polymerase Chain Reaction
Brain
Exons
Recombinant Proteins
Lipoxins
Liver
Microsomes
Liver
DNA
Complementary
Molecular Sequence Data
Biophysics
Biology
Transfection
Catalysis
Complementary DNA
Animals
Amino Acid Sequence
Cytochrome P450 Family 4
RNA
Messenger
Northern blot
Molecular Biology
Gene
Prostaglandins A
Messenger RNA
Base Sequence
Models
Genetic
Sequence Homology
Amino Acid
Intron
Blotting
Northern
Molecular biology
Introns
Rats
Mice
Inbred C57BL
Alternative Splicing
chemistry
5' Untranslated Regions
Laurates
Zdroj: Archives of Biochemistry and Biophysics. 383:225-232
ISSN: 0003-9861
DOI: 10.1006/abbi.2000.2078
Popis: We have isolated a cDNA for a mouse leukotriene B4 omega-hydroxylase, CYP4F14. The cDNA encoded a protein with 524 amino acids, whose sequence similarity is 95% that of rat CYP4F1. The microsomes from yeast cells transfected with CYP4F14 expression vector showed 0.1 nmol P450/mg protein and catalyzed omega-hydroxylations of leukotriene B4, 6-trans-leukotriene B4, lipoxin A4, prostaglandin A1, and several hydroxyeicosatetraeonic acids (HETEs), with 8-HETE being the most active substrate. In contrast, no activity was detected toward lipoxin B4, laurate, and arachidonate. The mRNA for CYP4F14 had three different 5' untranslated sequences. Analysis of the CYP4F14 gene showed that two exon I sequences with different transcription start sites are located in the gene, and two splicing signals on the 3' end of intron I are alternatively used. The mRNA for this P450 was detected only in the liver by Northern blot analysis, whereas a small amount of the mRNA was detected in the brain using RT-PCR. Administration of clofibrate had no effect on microsomal 6-trans-leukotriene B4 omega-hydroxylase activity, but resulted in a marked reduction in the content of mRNA for this P450 in the liver. These findings indicate that CYP4F14 is very similar to CYP4F1 except for its expression in the brain and 5' untranslated sequences.
Databáze: OpenAIRE
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