Mechanism(s) of activation of secretory phospholipase A(2)s in mouse keratinocytes.
The differential activation of different members of the phospholipase A(2) (PLA(2)) superfamily and their regulation are important as one or more of them regulates the production of eicosanoids and others may contribute to the formation of other lipid mediators. We previously reported the existence of two forms of secretory or sPLA(2) in mouse keratinocytes, namely type I and type II sPLA(2). We show here that mouse keratinocyte sPLA(2)s were potently activated by protease treatment and inhibited by protease inhibitors. We also observed that G protein effectors induced substantial release of oleic acid (OA) from prelabeled mouse keratinocytes. A G(i)/G(0) protein activator significantly enhanced the hydrolysis of OA and this increase was not responsive to either pertussis toxin or cholera toxin treatment. Although there was a significant negative correlation between intracellular cAMP levels and OA hydrolysis, experimentally increasing cAMP with forskolin treatment had no effect on sPLA(2) activity. Arachidonic acid but not its metabolites was also shown to marginally activate keratinocyte sPLA(2) by 1.5-fold. These results lead to the conclusion that mouse keratinocyte sPLA(2)s can be regulated primarily by proteolytic activation and a G protein pathway.
Li-Stiles
B
B
The University of Texas M. D. Anderson Cancer Center, Science Park-Research Division, P.O. Box 389, Smithville, TX 78957, USA.
Fischer
S M
SM
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism; D006868:Hydrolysis; D015603:Keratinocytes / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D051379:Mice; D019301:Oleic Acid / Q000378:metabolism; D010741:Phospholipases A / Q000145:classification / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D005576:Colforsin; D016718:Arachidonic Acid; D019301:Oleic Acid; D000242:Cyclic AMP; D010741:Phospholipases A; D014357:Trypsin; D019204:GTP-Binding Proteins