The proliferative properties and the ability to stimulate the Na+/H+antiport activity of a secretory phospholipase A2 were studied in rat aortic smooth muscle cells in culture. The requirement of the enzymatic activity of phospholipase A2 to elicit mitogenesis was assessed by the use of ammodytin L, a Ser49 phospholipase A2 from the venom of Vipera ammodytes, devoid of hydrolytic activity. We propose that the proliferative effect is mediated by the same transduction pathway for both proteins. In particular, 1) both secretory phospholipase A2 and ammodytin L stimulated thymidine incorporation in a dose-dependent manner; 2) both proteins affected the cell cycle, as assessed by cell growth and fluorescence-activated cell sorting experiments; 3) both phospholipase A2 and ammodytin L increased intracellular pH, a permissive factor for cell proliferation, through activation of the Na+/H+antiport; 4) ammodytin L was able to displace the 125I-labeled phospholipase A2 from specific binding sites in a concentration range consistent with that capable of eliciting a cellular response; and 5) the inhibition by heparin was similar for both proteins, taking into account the ratio of heparin to protein. In conclusion, the enzymatic activity of phospholipase A2 is not required for the stimulation of mitogenesis. The inhibitory effect of heparin combined with its therapeutic potential could help to clarify the role of phospholipase A2 in the pathogenesis of several preinflammatory situations.
Protein Dynamics Influence the Enzymatic Activity of Phospholipase A/Acyltransferases 3 and 4
