Continuous spectrophotometric assay of mammalian phosphoinositide-specific phospholipase Cdelta1 with a thiophosphate substrate analog.

Abstract excerpt

1,2-Dimyristoyloxypropane-3-thiophospho(1D-1-myo-inositol) (D-thio-DMPI) was used as a substrate for the continuous assay of phosphoinositide-specific phospholipase C (PI-PLC). Its activity with a Delta(1-132) deletion mutant of mammalian PI-PLCdelta1 is about one-fourth that with PI under similar conditions. Optimal conditions for the assay include 0.2 mM substrate, 0.2 mM Ca2+, and a mole ratio of...

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