Cloning and sequence analysis of the cDNA for arachidonate 12-lipoxygenase of porcine leukocytes.
The complete amino acid sequence of arachidonate 12-lipoxygenase (EC 1.13.11.31) of porcine leukocytes was deduced by cloning and sequence analysis of DNA complementary to its mRNA. The sequence was confirmed by automated Edman degradation of the N-terminal regions of the native enzyme and its proteolytic fragments. The cDNA had an open reading frame encoding 662 amino acid residues with a calculated molecular weight of 74,911. Amino acid residues 533-545, Cys-(Xaa)3-Cys-(Xaa)3-His-(Xaa)3-His, showed significant homology to the short cysteine- or histidine-containing sequences proposed as the metal-binding domains of transcription factors and various metal-containing proteins [Berg, J. M. (1986) Science 232, 485-487]. The amino acid sequence of 12-lipoxygenase exhibited 86% identity with human reticulocyte 15-lipoxygenase and showed 41% identity with human leukocyte 5-lipoxygenase. The 12-lipoxygenase cDNA recognized a 3.4-kilobase mRNA species in various porcine cell types, with the largest amount in leukocytes, followed by pituitary, lung, jejunum, and spleen.
Yoshimoto
T
T
Department of Biochemistry, Tokushima University School of Medicine, Japan.
Suzuki
H
H
Yamamoto
S
S
Takai
T
T
Yokoyama
C
C
Tanabe
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D001091:Arachidonate Lipoxygenases / Q000235:genetics*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA / Q000097:blood / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D007962:Leukocytes / Q000201:enzymology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D010446:Peptide Fragments / Q000302:isolation & purification; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013552:Swine
D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D004247:DNA; D001091:Arachidonate Lipoxygenases; D001092:Arachidonate 12-Lipoxygenase