Summary
Subtilisin is a bacterial serine protease with a broad specificity in the S1 subsite. It has been very extensively studied using a variety of kinetic and physical techniques. A chemical derivative, thiolsubtilisin, has been subjected to similar studies in order to analyze the effects of the OH to SH conversion on enzyme activity.
The native structure of thiosubtilisin is indicated by a variety of physical techniques. Oligopeptides bind nearly equally well to both enzymes, and a peptide chloromethylketone is much more reactive to thiolsubtilisin than to subtilisin. Both enzymes have a similar level of activity towards activated nonspecific amides and esters. However, thiolsubtilisin is inactive towards highly specific peptide amides and esters. Thiolsubtilisin also does not show good binding to boronic and arsonic acids. The observation that these transition state analog inhibitors bind poorly to thiolsubtilisin while other compounds bind nearly equally well to both enzymes suggests that thiolsubtilisin may not be able to stabilize the transition state during acylation by specific substrates.
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Abbreviations
- Ac X:
-
N-acetyl derivative of peptide X
- Ac SEt:
-
thioethyl acetate
- Ala(1-NPH):
-
beta (1-naphthyl)alanine
- Ala(2-NPH):
-
beta (2-naphthyl)alanine
- Ala(6-Qui):
-
beta (6-quinolyl)alanine
- Boc X:
-
N-tertiary-butyloxycarbonyl derivative of peptide X
- Bz X:
-
N-Benzoyl derivative of peptide X
- Bzc:
-
2-benzylcarbazate
- X CH2Cl:
-
chloromethylketone analog of amino acid X
- X CHO:
-
aldehyde analog of amino acid X
- Cys(Bzl):
-
S-benzyl derivative of cysteine
- Cys(DPM):
-
S-diphenylmethyl derivative of cysteine
- DMAC X:
-
4-dimethylaminocinnamic acid derivative of X
- DOPA:
-
3,4-dihydroxyphenylalanine
- EDTA:
-
ethylenediamine tetraacetic acid
- F3Ac (N-Me)pNA:
-
N-methyl, para-nitroanilide derivative of trifluoroacetic acid
- IND:
-
indole ring system
- Lim:
-
pH-independent limiting value
- Mec:
-
2-methylcarbazate
- McCN:
-
acetonitrile
- α-Naphth:
-
alpha naphthyl ring system
- X NH2 :
-
amide of peptide X
- X OET:
-
ethyl ester of peptide X
- X OMe:
-
methyl ester of peptide X
- X OpNP:
-
nitrophenyl ester of compound X
- P1, P2, etc.:
-
residue designation in an oligopeptide (32)
- Ph:
-
phenyl ring
- Phe(4-NHAc):
-
4-acetamido derivative of phenylalanine
- Phe(NO2):
-
4-nitrophenylalanine
- X pNA:
-
para-nitroanilide of compound X
- 2-PrOH:
-
2-propanol
- X SBz:
-
thiobenzyl ester of peptide X
- SHSTL:
-
thiolsubtilisin
- X-SHSTL:
-
thiolsubtilisin acylated by X
- SSI:
-
streptomyces subtilisin inhibitor
- STL:
-
subtilisin
- Tos X:
-
N-tosyl derivative of peptide X
- Trp(CHO):
-
N-formyl derivative of tryptophan
- Trp(NCps):
-
2-(2-nitro-4-carboxyphenylsulfenyl)-tryptophan
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Philipp, M., Bender, M.L. Kinetics of subtilisin and thiolsubtilisin. Mol Cell Biochem 51, 5–32 (1983). https://doi.org/10.1007/BF00215583
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DOI: https://doi.org/10.1007/BF00215583