title: Rhinovirus 3C protease catalyzes efficient cleavage of a fluorescein-labeled peptide affording a rapid and robust assay.
abstract: The 3C protease encoded by human rhinovirus type 2 catalyzes with equal efficiency cleavage of a peptide substrate with or without a fluorescein label attached to the amino acid at the P7' position. Substrates Ac-MEALFQGPLQYKDL-NH2 and MEALFQGPLQYKE(fluorescein)L are hydrolyzed with values of Vmax/KM of 970 M-1 s-1 and 1100 M-1 s-1, respectively. With the labeled substrate, HPLC achieves separation of substrate and product in 2.5 min. Separation in as little as 12 s is feasible. Fluorescein was derivatized so that it could be incorporated into peptides using automated solid-phase peptide synthesis.
authors.affiliation: Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, Connecticut 06877.
authors.forename: J L
authors.initials: JL
authors.lastname: Hopkins
authors.forename: R
authors.initials: R
authors.lastname: Betageri
authors.forename: K A
authors.initials: KA
authors.lastname: Cohen
authors.forename: M J
authors.initials: MJ
authors.lastname: Emmanuel
authors.forename: C R
authors.initials: CR
authors.lastname: Joseph
authors.forename: P M
authors.initials: PM
authors.lastname: Bax
authors.forename: P V
authors.initials: PV
authors.lastname: Pallai
authors.forename: M T
authors.initials: MT
authors.lastname: Skoog
mesh_terms: D000086042:3C Viral Proteases; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D005452:Fluoresceins; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D012229:Rhinovirus / Q000201:enzymology* / Q000235:genetics; D014764:Viral Proteins
chemical_list: D005452:Fluoresceins; D010455:Peptides; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases