Development of new synthetic methods for the preparation of phospholipids is one of the key steps in advancing membrane chemistry and biochemistry today. The compounds are needed for the study of biomembranes and membrane-bound enzymes to delineate phospholipid-phospholipid and phospholipid-protein interactions, as well as for mechanistic elucidation of phospholipid metabolizing enzymes. Specifically, phospholipases A2 (PLA2s) comprise a group of intracellular and secreted enzymes that catalyze the hydrolysis of the sn-2-ester bond of glycerophospholipids. The mammalian family of secreted phospholipases (sPLA2s) includes ten catalytically active isoforms. They have been implicated in a series of physiological and pathophysiological processes including lipid digestion, cell proliferation, neurosecretion, antibacterial defense, cancer, tissue injury, and atherosclerosis. In addition, individual phospholipase A2 enzymes play important and diverse roles in biological events by acting through multiple mechanisms: involving generation of lipid mediators, and executing their own action on their specific extracellular targets. Thus, availability of PLA2 directed fluorogenic substrates should enable to detect, and measure PLA2 catalysis, contribute to kinetic characterization and mechanistic elucidation of the enzymes and to better understand their exact physiological roles that are yet to be elucidated. We have designed a series of phospholipid analogues with chain-terminal reporter groups, including fluorescent donor-acceptor pairs, and fluorophore-quencher labels, with PLA1-resistant sn-1-substituents for PLA2 specific spectrophotofluorimetric assay of the enzyme. The synthesis relies a two-prong approach 1) using either a glyceric acid scaffold, or 2) based on a sequence using (R)-3-amino-1,2-propanediol as precursor of the PLA2 substrates. Details of the syntheses and formation of the hydrolysis products will be presented.

Design and synthesis of phospholipase A<sub>2 </sub>?directed fluorogenic substrates

248th National Meeting (2014)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.

Design and synthesis of phospholipase A2 ?directed fluorogenic substrates

Design and synthesis of phospholipase A₂?directed fluorogenic substrates