Molecular cloning and expression of human arachidonate 12-lipoxygenase

Biochem Biophys Res Commun. 1990 Nov 15;172(3):1230-5. doi: 10.1016/0006-291x(90)91580-l.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arachidonate 12-Lipoxygenase / biosynthesis
  • Arachidonate 12-Lipoxygenase / genetics*
  • Arachidonate 15-Lipoxygenase / genetics
  • Arachidonate 5-Lipoxygenase / genetics
  • Base Sequence
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • DNA / biosynthesis
  • DNA / chemistry
  • Dimethyl Sulfoxide / pharmacology
  • Gene Expression Regulation, Enzymologic*
  • Genomic Library
  • Humans
  • Leukemia, Erythroblastic, Acute / enzymology*
  • Molecular Sequence Data
  • Open Reading Frames
  • Restriction Mapping
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tumor Cells, Cultured / drug effects

Substances

  • DNA
  • Arachidonate 12-Lipoxygenase
  • Arachidonate 15-Lipoxygenase
  • Arachidonate 5-Lipoxygenase
  • Tetradecanoylphorbol Acetate
  • Dimethyl Sulfoxide

Associated data

  • GENBANK/M62982