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Literature summary for 3.4.17.18 extracted from

  • Grishin, A.M.; Akparov, V.K.h.; Chestukhina, G.G.
    Leu254 residue and calcium ions as new structural determinants of carboxypeptidase T substrate specificity (2008), Biochemistry (Moscow), 73, 1140-1145.
    View publication on PubMed
Search for more literature for 3.4.17.18

Cloned(Commentary)

Cloned (Comment) Organism
into the vector pET23a for expression in Escherichia coli BL21DE3 pLysS cells Thermoactinomyces vulgaris

Protein Variants

Protein Variants Comment Organism
H68N mutant, it is shown that the His68 residue is not a structural determinant of specificity Thermoactinomyces vulgaris
L254N mutant, hydrolysis efficiency of substrates with C-terminal Leu and Arg not changed, 28-fold decrease in activity towards the substrate with C-terminal Glu Thermoactinomyces vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
 N-benzyloxycarbonyl-Ala-Ala-Leu mutant H68N Thermoactinomyces vulgaris
0.046
-
 N-benzyloxycarbonyl-Ala-Ala-Leu wild-type enzyme Thermoactinomyces vulgaris
0.063
-
 N-benzyloxycarbonyl-Ala-Ala-Leu mutant L254N Thermoactinomyces vulgaris
4
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant L254N Thermoactinomyces vulgaris
4.4
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine wild-type enzyme Thermoactinomyces vulgaris
5.9
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant H68N Thermoactinomyces vulgaris

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+
-
 Thermoactinomyces vulgaris

Organism

Organism UniProt Comment Textmining
Thermoactinomyces vulgaris
-
-
-

Purification (Commentary)

Purification (Comment) Organism
by gel filtration on a Superdex-75 column and by using a CABS-Sepharose column Thermoactinomyces vulgaris

Renatured (Commentary)

Renatured (Comment) Organism
refolding after treatment with 8 M urea Thermoactinomyces vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dinitrophenyl-Ala-Ala-Arg + H2O
-
 Thermoactinomyces vulgaris 2,4-dinitrophenyl-Ala-Ala + Arg
-
 ?
N-2,4-dinitrophenyl-Ala-Ala-Arg + H2O
-
 Thermoactinomyces vulgaris N-2,4-dinitrophenyl-Ala-Ala-Arg + L-arginine
-
 ?
N-benzyloxycarbonyl-Ala-Ala-Glu + H2O
-
 Thermoactinomyces vulgaris N-benzyloxycarbonyl-Ala-Ala + L-glutamate
-
 ?
N-benzyloxycarbonyl-Ala-Ala-Leu + H2O
-
 Thermoactinomyces vulgaris N-benzyloxycarbonyl-Ala-Ala + L-leucine
-
 ?

Synonyms

Synonyms Comment Organism
carboxypeptidase T
-
 Thermoactinomyces vulgaris
CPT
-
 Thermoactinomyces vulgaris
metallocarboxypeptidase T
-
 Thermoactinomyces vulgaris

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 activity assay, N-benzyloxycarbonyl-Ala-Ala-Leu and N-benzyloxycarbonyl-Ala-Ala-Glu Thermoactinomyces vulgaris
37
-
 activity assay, N-2,4-dinitrophenyl-Ala-Ala-Arg Thermoactinomyces vulgaris

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.6
-
 N-benzyloxycarbonyl-Ala-Ala-Leu mutant L254N Thermoactinomyces vulgaris
8.4
-
 N-benzyloxycarbonyl-Ala-Ala-Leu mutant H68N Thermoactinomyces vulgaris
10
-
 N-benzyloxycarbonyl-Ala-Ala-Leu wild-type enzyme Thermoactinomyces vulgaris
14
-
 N-2,4-dinitrophenyl-Ala-Ala-Arg mutant L254N Thermoactinomyces vulgaris
15
-
 N-2,4-dinitrophenyl-Ala-Ala-Arg wild-type enzyme Thermoactinomyces vulgaris
18
-
 N-2,4-dinitrophenyl-Ala-Ala-Arg mutant H68N Thermoactinomyces vulgaris

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 activity assay Thermoactinomyces vulgaris