title: 6,8-Difluoro-4-methylumbiliferyl phosphate: a fluorogenic substrate for protein tyrosine phosphatases.
abstract: The fluorogenic substrate 6,8-difluoro-4-methylumbiliferyl phosphate (DIFMUP) has been widely used for the detection of serine and threonine phosphatase activities. Here we describe the use of this substrate for the characterization of protein tyrosine phosphatases (PTPs) and for the screening for PTP inhibitors. The measured kinetic and inhibitor constants for DIFMUP cleavage were comparable with those of the widely used but less discriminative and practicable substrates, para-nitrophenylphosphate and phosphotyrosine-containing peptides, respectively. Furthermore, the continuous and highly sensitive assay allows fast and accurate investigations of the type, kinetic behavior, and binding mode of small-molecule inhibitors. We discuss the validation of this assay system for various PTPs and its use in inhibitor screening for PTP1B.
authors.affiliation: Aventis Pharma Deutschland GmbH--A Company of Sanofi-Aventis, TD Metabolism, 65926 Frankfurt am Main, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Welte
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Baringhaus
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schmider
authors.forename: Günter
authors.initials: G
authors.lastname: Müller
authors.forename: Stefan
authors.initials: S
authors.lastname: Petry
authors.forename: Norbert
authors.initials: N
authors.lastname: Tennagels
mesh_terms: D004353:Drug Evaluation, Preclinical; D005456:Fluorescent Dyes / Q000378:metabolism*; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism*; D007700:Kinetics; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: C119285:6,8-difluoro-4-methylumbelliferyl phosphate; D005456:Fluorescent Dyes; D006923:Hymecromone; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases