PLA2

Cytosolic phospholipase A2, cPLA2, is an important regulator of eicosanoid biosynthesis. Its role has been implemented in a variety of inflammatory diseases. Recently, ceramide and ceramide-1-phosphate have been identified as stimulators of cPLA2 activity.

Aims of this study:

• To verify that ceramide and ceramide-1-phosphate induce translocation of cPLA2 from the cytosol to intracellular membranes where it is activated
• To identify amino acid residues responsible for cPLA2 – ceramide interactions

The confocal laser scanning microscope is being used to follow intracellular movements of cPLA2 as a response to ceramide stimuli. Fusion proteins have been constructed where cPLA2 or its sub domains have been coupled to fluorescent protein tags, such as EGFP and dsRED. A number of structural mutants have also been constructed to determine the residues responsible for chemical interactions and cPLA2 translocation.