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

Journal Article · Sat Jun 01 04:00:00 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), Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Vol. 5:9; ISSN FAJOE; ISSN 0892-6638
• Country of Publication: United States
• Language: English

Related Subjects

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