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Updated: Aug 15, 2026

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes
Published on: May 25, 2018
Spectral titration of active site of carboxypeptidase A
Abstract:
In the carboxypeptidase A-catalyzed hydrolysis of O-[trans-alpha-(benzoylamino)-cinnamoyl]-L-beta-phenyllactate (BACPL) or O-[trans-alpha-(benzoylamino)-p-(phenylazo)cinnamoyl]-L-beta-phenyllacta te (BAPACPL), biphasic kinetic behavior was observed due to the accumulation of an intermediate. At -12 degrees C, conversion of the intermediate into the product was much slower than the formation of the intermediate, which accumulated in quantitative amounts. From the absorbance changes observed during the formation process of the intermediate, the concentration of active site of the enzyme was determined.
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