A Real-Time Fluorogenic Phospholipase A2 Assay for Biochemical and Cellular Activity Measurements - ScienceDirect
摘要：
A fluorogenic analog of the PLA 2 substrate PC, named Dabcyl-BODIPY-PC, or simply DBPC, was synthesized with a fluorescence quencher (Dabcyl, 4-[(4-[ N,N-dimethylamino]phenyl)azo]benzoic acid) in the sn-1 acyl chain and a BODIPY fluor in the sn-2 acyl chain. DBPC was recognized by sPLA 2 from each of the four sources examined (bee venom, human synovial fluid, cobra venom, and bovine pancreas). A dramatic and quantifiable fluorescence enhancement of DBPC occurred upon phospholipase digestion both in the presence and absence of excess PC. Both real-time and endpoint assays for PLA 2 were sensitive, consistent, and rapid. Thus, DBPC can be used as a sensitive fluorogenic probe for in vitro high-throughput screening assays for PLA 2 activation and inhibition and would expedite studies of PLA 2 in cellular signaling, in vitro screening for drug discovery, and subcellular localization of enzyme activity.
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关键词：
Animals Cattle Dogs Humans Synovial Fluid Pancreas Epithelial Cells Boron Compounds Phospholipases Phospholipases A
DOI：
10.1016/S1074-5521(02)00168-0
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年份：
2002
Elsevier
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