Type II phospholipase A2 recombinant overexpression enhances stimulated arachidonic acid release.
The coding sequence of type II phospholipase A2 from human placenta was cloned in a bovine papilloma virus-derived eukaryotic expression vector under the control of the metallothionein promoter. Stably transfected C127 mouse fibroblast lines were obtained with this vector. These transfected cells overexpressed a functional 14 kDa phospholipase A2, which was bulky secreted. However, a significant phospholipase A2 activity was measured in cell homogenates. The involvement of this 14 kDa phospholipase A2 in mechanisms related to stimulated arachidonic acid release was investigated. We could parallel the overexpression of phospholipase A2 with an increase in phorbol ester and fluoroaluminate-stimulated arachidonic acid release. Pertussis toxin inhibited this stimulation. These results suggest that the 14 kDa type II phospholipase A2 might contribute to stimulation of arachidonic acid release, and therefore to eicosanoid production.
Pernas
P
P
URA CNRS 1283, Laboratoire de Biochimie CHU Saint Antoine, Université Pierre et Marie Curie, Paris, France.
Masliah
J
J
Olivier
J L
JL
Salvat
C
C
Rybkine
T
T
Bereziat
G
G
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D037342:Pertussis Toxin; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*; D054467:Phospholipases A2; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D001095:Arachidonic Acids; D007527:Isoenzymes; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin; D010741:Phospholipases A; D054467:Phospholipases A2