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

Abstract

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.

Authors:
; ; ; ;  [1]
  1. Vanderbilt Univ., Nashville, TN (United States)
Publication Date:
OSTI Identifier:
5822913
Resource Type:
Journal Article
Journal Name:
FASEB Journal (Federation of American Societies for Experimental Biology); (United States)
Additional Journal Information:
Journal Volume: 5:9; Journal ID: ISSN 0892-6638
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Funk, C D, Funk, L B, Kennedy, M E, Pong, A S, and Fitzgerald, G A. Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment. United States: N. p., 1991. Web.
Funk, C D, Funk, L B, Kennedy, M E, Pong, A S, & Fitzgerald, G A. Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment. United States.
Funk, C D, Funk, L B, Kennedy, M E, Pong, A S, and Fitzgerald, G A. 1991. "Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment". United States.
@article{osti_5822913,
title = {Human platelet/erythroleukemia cell prostaglandin G/H synthase: cDNA cloning, expression, and gene chromosomal assignment},
author = {Funk, C D and Funk, L B and Kennedy, M E and Pong, A S and Fitzgerald, G A},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/5822913}, journal = {FASEB Journal (Federation of American Societies for Experimental Biology); (United States)},
issn = {0892-6638},
number = ,
volume = 5:9,
place = {United States},
year = {Sat Jun 01 00:00:00 EDT 1991},
month = {Sat Jun 01 00:00:00 EDT 1991}
}