Fluorogenic peptide substrates for serine and threonine phosphatases.
A new fluorescent assay for Ser/Thr protein phosphatases has been developed. Hydrolysis of a phosphoSer residue liberates the Ser hydroxyl group, which induces a cyclization reaction on the N-terminal carbamate and releases a fluorescent reporter. Sequence selectivity is observed using several peptide substrates against alkaline phosphatase (ALP), bacteriophage lambda protein phosphatase (lambda-PPase), and vaccinia H1 related phosphatase (VHR). These studies suggest that the assay could be a useful tool for profiling the substrate specificities of medicinally important phosphatases.
Xue
Fengtian
F
Department of Chemistry, Brown University, Providence, Rhode Island 02912, USA.
Seto
Christopher T
CT
D005456:Fluorescent Dyes / Q000737:chemistry*; D008958:Models, Molecular; D010455:Peptides / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D010455:Peptides; D010749:Phosphoprotein Phosphatases