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P2052
Selective Detection of Pyrophosphate Using a Fluorogenic Pyrophosphate Sensor
Presenter Jianjun He, ABD Bioquest, Inc., USA
Additional Authors: Jinfang Liao, Jianjun He, Chunmei Wei, and Zhenjun Diwu
Pyrophosphate (PPi) are produced by a number of biochemical reactions, such as ATP hydrolysis, DNA and RNA polymerizations, cyclic AMP formation by the enzyme adenylate cyclase, and the enzymatic activation of fatty acids to form their coenzyme A esters. Many pathological disorders such as familial chondrocalcinosis are associated with abnormal regulation of the transport of inorganic pyrophosphate. Our PhosphoWorks™ PP Lite Blue-based pyrophosphate detection system has been developed to provide the most robust spectrophotometric method for measuring pyrophosphate. PP Lite Blue is a fluorogenic pyrophosphate sensor that has its fluorescence intensity proportionally dependent upon the concentration of pyrophosphate. The PP Lite Blue-based pyrophosphate assay is much more robust than the enzyme-coupling pyrophosphate methods that require at least two enzymes for pyrophosphate detection. The enzyme-coupling pyrophosphate methods often give high rate of false positive hits due to the use of multiple enzymes that are inhibited by some library compounds. The PP Lite Blue-based PPi assay can be performed in a convenient 384-well or 1536-well microtiter-plate format, and can be easily adapted to automation with no separation steps required.