Abstract
The synthetic peptide Chymohelizyme-1 (CHZ-1) exhibits esterase activity against carbobenzoxytyrosinep-nitrophenyl ester (ZTONP), carbobenzoxyalaninep-nitrophenyl ester (ZAONP), andt-butyloxycarbonyltyrosinep-nitrophenyl ester (BocTONP). However, earlier reports of catalytic activity against less labile esters and amides have proven to be incorrect. The major reason for the errors appears to have been the omission of certain controls in the previous work. Although the catalytic triad does not appear to be functioning as designed, the catalytic activity of CHZ-1 does depend on the integrity of its primary structure. The pH dependence of hydrolysis of ZTONP points to general-base catalysis, whereas a preference for hydrophobic substrates suggest that the structure of CHZ-1 is performing some other role in assisting catalysis.
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Abbreviations
- ADH:
-
alcohol dehydrogenase
- ATEE:
-
acetyltyrosine ethyl ester
- BocTONP:
-
t-butyloxycarbonyltyrosinep-nitrophenyl ester
- BTEE:
-
benzoyltyrosine ethyl ester
- CHES:
-
2-[N-cyclohexylamino]-ethanesulfonic acid
- CHZ-1:
-
Chymohelizyme-1 (blocking group on histidine α-amino group is acetyl)
- CHZ-1A:
-
chymohelizyme-1A (blocking group on histidine α-amino group ist-butylacetyl)
- HPLC:
-
high-performance liquid chromatography
- NAD+ :
-
nicotinamide adenine dinucleotide
- ONP:
-
p-Nitrophenyl ester
- PMSF:
-
phenylmethylsulfonyl fluoride
- SBTI:
-
soybean trypsin inhibitor
- TFA:
-
trifluoroacetic acid
- TPCK:
-
tosylphenylalanine chloromethyl ketone
- ZAONP:
-
benzyloxycarbonylalaninep-nitrophenyl ester
- ZTONP:
-
benzyloxycarbonyltyrosinep-nitrophenyl ester
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Corey, M.J., Hallakova, E., Pugh, K. et al. Studies on chymotrypsin-like catalysis by synthetic peptides. Appl Biochem Biotechnol 47, 199–212 (1994). https://doi.org/10.1007/BF02787935
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DOI: https://doi.org/10.1007/BF02787935