Synthesis of Fluorogenic Substrates for Continuous Assay of Phosphatidylinositol-Specific Phospholipase C

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作者：

TO Zaikova，AV Rukavishnikov，GB Birrell，OH Griffith，JFW Keana

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摘要：

An improved synthesis of fluorogenic substrate analogues for phosphatidylinositol-specific phospholipase C (PI-PLC) is described. The water-soluble substrates, which are derived from fluorescein, are not fluorescent until cleaved by the enzyme, and provide a convenient means to continuously monitor PI-PLC activity. The improvement in the synthesis lies in the method used to protect the hydroxyl groups of the inositol portion of the substrate molecule and allows a milder deprotection procedure to be used. The result is a much more reproducible synthesis of the substrate. The improved procedure has been employed to synthesize a series of fluorogenic substrates, which differ in the length of the aliphatic tail attached to the fluorescein portion of the molecule. The length of the tail was found to have a significant effect on the rate of cleavage of these substrates.

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关键词：

Fluorescein Phosphatidylinositol Diacylglycerol-Lyase Fluorescent Dyes Molecular Structure Type C Phospholipases Phosphoinositide Phospholipase C

DOI：

10.1177/0165551508091309

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年份：

2001