Manna_2007_Methods.Enzymol_434_15

Phospholipase A(2)s (PLA(2)s) are a superfamily of enzymes involved in production of a wide variety of lipid mediators, including arachidonic acid, lysophospholipids, platelet activation factor, and eicosanoids. Fluorescence-based, real-time cellular activity assays for PLA(2)s have been developed as a tool for studying the function and spatiotemporal regulation of PLA(2)s. Recent progress in fluorogenic phospholipid design, genetic methods, and multiphoton multichannel microscopy allows simultaneous and continuous measurement of cellular localization and activity of PLA(2) in an isoform-selective manner. These assays should aid in elucidating the physiological roles and regulatory mechanisms of diverse PLA(2) isoforms, developing isoform-specific PLA(2) inhibitors, and analyzing lipidomics data of PLA(2) products.