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J. Biol. Chem., Vol. 269, Issue 4, 2365-2368, 01, 1994
J Chen, SJ Engle, JJ Seilhamer and JA Tischfield
Extensive biochemical studies of phospholipase A2s (PLA2s) over the last
two decades indicate that there are likely to be several distinct PLA2
genes in mammals. Here we report the cloning of a 1-kilobase pair cDNA
encoding a novel human low molecular weight PLA2. The cDNA appears to
encode a 118-amino acid mature peptide (M(r) = 13,592) preceded by a
20-residue prepeptide. The deduced amino acid sequence encodes a protein
that lacks one of the seven disulfide bridges found in similar PLA2s and,
therefore, represents a class of enzymes distinct from the mammalian group
I and group II enzymes. An RNA blot hybridized with the cDNA exhibited a
putative 1.2-kilobase pair transcript in heart and, less abundantly, in
lung, as well as multiple putative transcripts in placenta. When the cDNA
was expressed using an Epstein-Barr virus-based vector in human 293s cells,
PLA2 activity accumulated in the culture medium. Conditioned medium
optimally hydrolyzed the phospholipids of [1- 14C]oleate-labeled
Escherichia coli at neutral to alkaline pH with 10 mM or greater Ca2+. In
assays done with individual substrates, L-alpha- 1-palmitoyl-2-oleoyl
phosphatidylcholine was more efficiently hydrolyzed than
L-alpha-1-palmitoyl-2-arachidonyl phosphatidylcholine,
L-alpha-1-palmitoyl-2-arachidonyl phosphatidylethanolamine, or L-alpha-
1-stearoyl-2-arachidonyl phosphatidylinositol.
Cloning and recombinant expression of a novel human low molecular weight Ca(2+)-dependent phospholipase A2
Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis 46202.
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