ASSAY AND SUBSTRATE FOR ARACHIDONOYL-SPECIFIC PHOSPHOLIPASE A2

摘要

A non-radioactive, spectrophotometric, microtiter plate assay for human cystolic phospholipase A (cPLA) is disclosed. The assay utilizes a novel synthetic thiolphospholipid analog as a substrate. In one embodiment, the substrate is a phosphatidylcholine derivative with an arachidonoylthioester in the sn-2 position and an alkyl-ether or alkenyl-ether in the sn-1 position. The alkyl-ether or alkenyl-ether in the sn-1 position of the substrate ensures that the assay will only measure PLA activity and will not be complicated by metabolism of the lysophospholipid product by the enzyme's lysophospholipase activity. As a model of the invention, the figure shows the 8 steps used to synthesize the compound 1-Hexadecyl-2-arachidonylthio-2-deoxy-sn-glycero-3-phosphorylcholine from (R)-glycidol.