Cloning of the cDNA for human 12-lipoxygenase

Journal Article · Mon Oct 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.19.7477· OSTI ID:5800900

Izumi, T; Hoshiko, S; Radmark, O; Samuelsson, B ^[1]

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1. Karolinska Institutet, Stockholm (Sweden)

A full-length cDNA clone encoding 12-lipoxygenase was isolated from a human platelet cDNA library by using a cDNA for human reticulocyte 15-lipoxygenase as probe for the initial screening. The cDNA had an open reading frame encoding 662 amino acid residues with a calculated molecular weight of 75,590. Three independent clones revealed minor heterogeneities in their DNA sequences. Thus, in three positions of the deduced amino acid sequence, there is a choice between two different amino acids. The deduced sequence from the clone plT3 showed 65{percent} identity with human reticulocyte 15-lipoxygenase and 42{percent} identity with human leukocyte 5-lipoxygenase. The 12-lipoxygenase cDNA recognized a 3.0-kilobase mRNA species in platelets and human erythroleukemia cells (HEL cells). Phorbol 12-tetradecanoyl 13-acetate induced megakaryocytic differentiation of HEL cells and 12-lipoxygenase activity and increased mRNA for 12-lipoxygenase. The identity of the cloned 12-lipoxygenase was assured by expression in a mammalian cell line (COS cells). Human platelet 12-lipoxygenase has been difficult to purify to homogeneity. The cloning of this cDNA will increase the possibilities to elucidate the structure and function of this enzyme.

• OSTI ID: 5800900
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:19; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

551001* -- Physiological Systems-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CARCINOGENS
CLONING
DNA
DNA HYBRIDIZATION
DNA-CLONING
ENZYMES
ESTERS
FUNCTIONS
HYBRIDIZATION
MAMMALS
MAN
MATERIALS
MESSENGER-RNA
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PHORBOL ESTERS
PRIMATES
PROMOTERS
RECOMBINANT DNA
RNA
TUMOR PROMOTERS
VERTEBRATES