Thiobenzyl benzyloxycarbonyl-L-lysinate, substrate for a sensitive colorimetric assay for trypsin-like enzymes
Thiobenzyl benzyloxycarbonyl-L-lysinate (Z-Lys-SBzl) a substrate for trypsin-like proteases, was synthesized. In the presence of 5,5'-dithiobis(2-nitrobenzoic acid) the hydrolysis of the thiol ester by trypsin-like enzymes provides a continuous colorimetric assay with a sensitivity comparable to the best fluorometric substrates. Z-Lys-SBzl is readily synthesized in good yield, is water soluble, and has a low rate of spontaneous hydrolysis even at pH 8.0. This assay procedure has been routinely used with urokinase, human urinary and human plasma kallikrein, thrombin, plasmin, ..beta..-trypsin, factor Xa, and crotalase. Levels of detection of these enzymes are in the range 10/sup -14/ to 10/sup -13/ mol.
- Research Organization:
- Brookhaven National Lab., Upton, NY
- OSTI ID:
- 6288931
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 93:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ABSORPTION SPECTROSCOPY
BIOASSAY
CHEMICAL REACTIONS
DATA
DATA FORMS
DECOMPOSITION
ENZYMES
EXPERIMENTAL DATA
HYDROLASES
HYDROLYSIS
INFORMATION
ISOLATED VALUES
LYSIS
NUMERICAL DATA
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDE HYDROLASES
SENSITIVITY
SOLVOLYSIS
SPECTROSCOPY
SUBSTRATES