Molecular cloning and expression of human arachidonate 12-lipoxygenase.
The cDNA for a 12-lipoxygenase was isolated from cDNA library of human erythroleukemia cells. The cDNA had an open reading frame encoding 663 amino acids with a calculated molecular weight of 75,513. The deduced amino acid sequence of human 12-lipoxygenase exhibited 41.5%, 65.3% and 65.4% identity with human 5-lipoxygenase, human 15-lipoxygenase and porcine 12-lipoxygenase, respectively. Blot hybridization analysis of RNA from human erythroleukemia cells demonstrated a single species (3.1 kb) of mRNA with the cDNA probe for 12-lipoxygenase of these cells, but not with the cDNA for porcine leukocyte enzyme. The cytosol of Escherichia coli transformed with a recombinant pUC19 plasmid oxygenated the position 12 of arachidonic acid.
Yoshimoto
T
T
Department of Biochemistry, Tokushima University School of Medicine, Japan.
Yamamoto
Y
Y
Arakawa
T
T
Suzuki
H
H
Yamamoto
S
S
Yokoyama
C
C
Tanabe
T
T
Toh
H
H
D000595:Amino Acid Sequence; D001092:Arachidonate 12-Lipoxygenase / Q000096:biosynthesis / Q000235:genetics*; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis / Q000737:chemistry; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D015698:Genomic Library; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D004247:DNA; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase; D001094:Arachidonate 5-Lipoxygenase; D013755:Tetradecanoylphorbol Acetate; D004121:Dimethyl Sulfoxide