title: Synthesis of fluorogenic substrates for continuous assay of phosphatidylinositol-specific phospholipase C.
abstract: 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.
authors: Department of Chemistry and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
authors: T O
authors: TO
authors: Zaikova
authors: A V
authors: AV
authors: Rukavishnikov
authors: G B
authors: GB
authors: Birrell
authors: O H
authors: OH
authors: Griffith
authors: J F
authors: JF
authors: Keana
mesh_terms: D019793:Fluorescein / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D019793:Fluorescein