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), Vol. 87:19; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

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