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Title: Molecular cloning and amino acid sequence of human 5-lipoxygenase

5-Lipoxygenase (EC 1.13.11.34), a Ca/sup 2 +/- and ATP-requiring enzyme, catalyzes the first two steps in the biosynthesis of the peptidoleukotrienes and the chemotactic factor leukotriene B/sub 4/. A cDNA clone corresponding to 5-lipoxygenase was isolated from a human lung lambda gt11 expression library by immunoscreening with a polyclonal antibody. Additional clones from a human placenta lambda gt11 cDNA library were obtained by plaque hybridization with the /sup 32/P-labeled lung cDNA clone. Sequence data obtained from several overlapping clones indicate that the composite DNAs 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 A/sub 4/ hydrolase or Ca/sup 2 +/-binding proteins. RNA blot analysis indicated substantial amounts of an mRNA species of approx. = 2700 nucleotides in leukocytes, lung, and placenta.
Authors:
; ; ; ; ;
Publication Date:
OSTI Identifier:
6464728
Resource Type:
Journal Article
Resource Relation:
Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States); Journal Volume: 85:1
Research Org:
Karolinska Institutet, Stockholm (Sweden)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; OXYGENASES; AMINO ACID SEQUENCE; RECOMBINANT DNA; DNA-CLONING; HYBRIDIZATION; INFLAMMATION; LUNGS; MAN; PHOSPHORUS 32; PLACENTA; ANIMALS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BODY; CLONING; DAYS LIVING RADIOISOTOPES; DNA; DNA HYBRIDIZATION; ENZYMES; FETAL MEMBRANES; ISOTOPES; LIGHT NUCLEI; MAMMALS; MEMBRANES; MOLECULAR STRUCTURE; NUCLEI; NUCLEIC ACIDS; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; ORGANS; OXIDOREDUCTASES; PATHOLOGICAL CHANGES; PHOSPHORUS ISOTOPES; PRIMATES; RADIOISOTOPES; RESPIRATORY SYSTEM; SYMPTOMS; VERTEBRATES 550201* -- Biochemistry-- Tracer Techniques