Molecular Cloning and Amino Acid Sequence of Human 5-lipoxygenase

• Published in: Proceedings of the National Academy of Sciences - PNAS 1988-01, Vol.85 (1), p.26-30
• Main Authors: Matsumoto, Takashi, Funk, Colin D., Rådmark, Olof, Höög, Jan-Olov, Jörnvall, Hans, Samuelsson, Bengt
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Amino Acids - analysis; Antibodies; Arachidonate 5-Lipoxygenase - genetics; Arachidonate Lipoxygenases - genetics; Base Sequence; Biological and medical sciences; Biotechnology; Cloning, Molecular; Complementary DNA; DNA - genetics; DNA Restriction Enzymes; Enzymes; Female; Fundamental and applied biological sciences. Psychology; Gels; Genes; Genetic engineering; Genetic technics; Gin; Humans; Leukocytes; Lung - enzymology; Lungs; Methods. Procedures. Technologies; Molecular cloning; Molecular Sequence Data; Nucleotide Mapping; Placenta; Placenta - enzymology; RNA; RNA, Messenger - genetics
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.85.1.26

5-Lipoxygenase (EC 1.13.11.34), a Ca2+- and ATP-requiring enzyme, catalyzes the first two steps in the biosynthesis of the peptidoleukotrienes and the chemotactic factor leukotriene B4. A cDNA clone corresponding to 5-lipoxygenase was isolated from a human lung λ gt11 expression library by immunoscreening with a polyclonal antibody. Additional clones from a human placenta λ gt11 cDNA library were obtained by plaque hybridization with the 32P-labeled lung cDNA clone. Sequence data obtained from several overlapping clones indicate that the composite cDNAs contain the complete coding region for the enzyme. From the deduced primary structure, 5-lipoxygenase encodes a 673 amino acid protein with a calculated molecular weight of 77,839. Direct analysis of the native protein and its proteolytic fragments confirmed the deduced composition, the amino-terminal amino acid sequence, and the structure of many internal segments. 5-Lipoxygenase has no apparent sequence homology with leukotriene A4 hydrolase or Ca2+-binding proteins. RNA blot analysis indicated substantial amounts of an mRNA species of ≈ 2700 nucleotides in leukocytes, lung, and placenta.