Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment

Journal Article · Sat Jun 01 00:00:00 EDT 1991 · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)

OSTI ID:5822913

Funk, C D; Funk, L B; Kennedy, M E; Pong, A S; Fitzgerald, G A ^[1]

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1. Vanderbilt Univ., Nashville, TN (United States)

Platelets metabolize arachidonic acid to thromboxane A{sub 2}, a potent platelet aggregator and vasoconstrictor compound. The first step of this transformation is catalyzed by prostaglandin (PG) G/H synthase, a target site for nonsteroidal antiinflammatory drugs. We have isolated the cDNA for both human platelet and human erythroleukemia cell PGG/H synthase using the polymerase chain reaction and conventional screening procedures. The cDNA encoding the full-length protein was expressed in COS-M6 cells. Microsomal fractions from transfected cells produced prostaglandin endoperoxide derived products which were inhibited by indomethacin and aspirin. Mutagenesis of the serine residue at position 529, the putative aspirin acetylation site, to an asparagine reduced cyclooxygenase activity to barely detectable levels, an effect observed previously with the expressed sheep vesicular gland enzyme. Platelet-derived growth factor and phorbol ester differentially regulated the expression of PGG/H synthase mRNA levels in the megakaryocytic/platelet-like HEL cell line. The PGG/H synthase gene was assigned to chromosome 9 by analysis of a human-hamster somatic hybrid DNA panel. The availability of platelet PGG/H synthase cDNA should enhance our understanding of the important structure/function domains of this protein and it gene regulation.

• OSTI ID: 5822913
• Journal Information: FASEB Journal (Federation of American Societies for Experimental Biology); (United States), Vol. 5:9; ISSN 0892-6638
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
BLOOD PLATELETS
MOLECULAR BIOLOGY
OXIDOREDUCTASES
GENES
ACETYLSALICYLIC ACID
DNA SEQUENCING
GENE REGULATION
GENETIC MAPPING
GROWTH FACTORS
MAN
PHORBOL ESTERS
PROSTAGLANDINS
STRUCTURE-ACTIVITY RELATIONSHIPS
TUMOR CELLS
ANALGESICS
ANIMAL CELLS
ANIMALS
ANTIPYRETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CARCINOGENS
CENTRAL NERVOUS SYSTEM DEPRESSANTS
DRUGS
ENZYMES
ESTERS
HYDROXY ACIDS
MAMMALS
MAPPING
MATERIALS
MITOGENS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
550200* - Biochemistry
550400 - Genetics