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New intramolecularly quenched fluorogenic peptide substrates for the study of the kinetic specificity of papain
Author(s) -
Garcia-Echeverria Carlos,
Rich Daniel H.
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80336-f
Subject(s) - chemistry , papain , hydrolysis , förster resonance energy transfer , cysteine , fluorescence , quenching (fluorescence) , benzoic acid , peptide , catalysis , acceptor , amino acid , stereochemistry , enzyme , organic chemistry , biochemistry , physics , quantum mechanics , condensed matter physics
A series of new substrates for determining the catalytic activity of cysteine proteinases is described. The rate of hydrolysis by papain was monitored by a fluorescence continuous assay based on internal resonance energy transfer using 5‐[(2‐aminoethyl)amino]naphtalene‐1‐sulfonic acid (EDANS) and 4‐(4‐dimethylaminophenylazo)benzoic acid (DABCYL) as fluorescent donor and quenching acceptor, respectively, in peptides with the general structure: DABCYL‐Lys‐Phe‐Gly‐Xxx‐Ala‐Ala‐EDANS. The substrates were used to evaluate the effect of amino acid structure in the Sl 1 position on the kinetic parameters for papain catalyzed hydrolysis.