An improved spectrophotometric assay for leucine aminopeptidase.
A sensitive assay to determine the activity of leucine aminopeptidase (EC 3.4.11.1), using L-leucine thiobenzyl ester as substrate, was developed. Hydrolysis of the ester by leucine aminopeptidase can be monitored in the presence of 5,5-dithiobis-(2-nitrobenzoic acid) by continuous spectrophotometric measurement at 412 nm. Comparison with some amide substrates showed that the thiol ester provides a much more sensitive assay, its specificity constant (Vmax./Km) being some 3000-fold higher than that of leucine p-nitroanilide.
D000818:Animals; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives; D007931:Leucyl Aminopeptidase / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods*; D013379:Substrate Specificity; D013552:Swine
C052813:leucine thiobenzyl ester; D007931:Leucyl Aminopeptidase; D007930:Leucine