Contamination of commercial preparations of calmodulin by phospholipase A2.
In the course of studies of the possible regulation of cellular phospholipase A2 activities by calcium and calmodulin, it was observed that some of the commercial preparations of calmodulin contained significant phospholipase A2 activity. Six commercially available calmodulin sources were compared for the presence of contaminating phospholipase A2 activity, relative purity by SDS-gel electrophoresis, and relative biological activity in stimulating calmodulin-deficient phosphodiesterase. One of the commercial calmodulin sources contained a relatively high specific phospholipase A2 activity (1.30 +/- 0.11 nmol [1-14C]arachidonic acid released/mg protein per h) and yielded two major bands in SDS-gel electrophoresis. Two of the calmodulin sources tested were relatively free of phospholipase A2 activity, were quite pure (one band on SDS-gel) and had high biological activity in stimulating calmodulin-deficient phosphodiesterase. Thus, investigators using commercially available preparations of calmodulin should be aware of the contamination of some of these sources by phospholipase A2 activity. These findings may be of importance to investigators considering the role of calmodulin in activating a variety of calcium-dependent enzymes, including phospholipase A2.
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000032:analysis; D001921:Brain / Q000378:metabolism; D002147:Calmodulin / Q000032:analysis; D002417:Cattle; D004340:Drug Contamination; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010740:Phospholipases / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D051381:Rats; D013552:Swine
D001095:Arachidonic Acids; D002147:Calmodulin; D016718:Arachidonic Acid; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases