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Molecular cloning and amino acid sequence of human 5-lipoxygenase

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
DOI:https://doi.org/10.1073/pnas.85.1.26· OSTI ID:6464728
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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.
Research Organization:
Karolinska Institutet, Stockholm (Sweden)
OSTI ID:
6464728
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 85:1; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DNA-CLONING
ENZYMES
FETAL MEMBRANES
HYBRIDIZATION
INFLAMMATION
ISOTOPES
LIGHT NUCLEI
LUNGS
MAMMALS
MAN
MEMBRANES
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
OXIDOREDUCTASES
OXYGENASES
PATHOLOGICAL CHANGES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLACENTA
PRIMATES
RADIOISOTOPES
RECOMBINANT DNA
RESPIRATORY SYSTEM
SYMPTOMS
VERTEBRATES