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Literature summary for 3.4.17.18 extracted from

  • Akparov, V.K.h.; Grishin, A.M.; Yusupova, M.P.; Ivanova, N.M.; Chestukhina, G.G.
    Structural principles of the wide substrate specificity of Thermoactinomyces vulgaris carboxypeptidase T. Reconstruction of the carboxypeptidase B primary specificity pocket (2007), Biochemistry (Moscow), 72, 416-423.
    View publication on PubMed
Search for more literature for 3.4.17.18

Application

Application Comment Organism
degradation reconstruction of the primary specificity pocket of CpT to make it like CpB neither enhances the CpT5 activity with a substrate possessing C-terminal Arg, nor lowers the activity with a substrate carrying C-terminal Leu. Notwithstanding the considerable structural similarity of CpT and CpB, the mechanisms underlying their substrate specificities are different Thermoactinomyces vulgaris

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutant pro-cpT genes cloned into pET23a vector and expressed in the Escherichia coli BL21(DE3)pLysS cells Thermoactinomyces vulgaris

Protein Variants

Protein Variants Comment Organism
A243G successive substitution of residues in the CpT S1'-subsite by similar residues of CpB, retains substrate specificity of the wild-type Thermoactinomyces vulgaris
A243G/D253G/T255D successive substitution of residues in the CpT S1'-subsite by similar residues of CpB, retains substrate specificity of the wild-type Thermoactinomyces vulgaris
D253G/T255D successive substitution of residues in the CpT S1'-subsite by similar residues of CpB, retains substrate specificity of the wild-type Thermoactinomyces vulgaris
G207S/A243G/D253G/T255D successive substitution of residues in the CpT S1'-subsite by similar residues of CpB, retains substrate specificity of the wild-type Thermoactinomyces vulgaris
G207S/A243G/T250A/D253G/T255D successive substitution of residues in the CpT S1'-subsite by similar residues of CpB, retains substrate specificity of the wild-type Thermoactinomyces vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant A243G Thermoactinomyces vulgaris
0.03
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant A243G/D253G/T255D Thermoactinomyces vulgaris
0.04
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant D253G/T255D Thermoactinomyces vulgaris
0.09
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant G207S/A243G/D253G/T255D Thermoactinomyces vulgaris
0.15
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant G207S/A243G/T250A/D253G/T255D Thermoactinomyces vulgaris
3.6
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant A243G Thermoactinomyces vulgaris
3.6
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant D253G/T255D Thermoactinomyces vulgaris

Organism

Organism UniProt Comment Textmining
Thermoactinomyces vulgaris P29068
-
-

Purification (Commentary)

Purification (Comment) Organism
native CpT isolated from inclusion bodies, purified by molecular exclusion chromatography and on cation-exchange resin Thermoactinomyces vulgaris

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
CPT
-
 Thermoactinomyces vulgaris

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2 8 benzyloxycarbonyl-alanyl-alanyl-leucine mutant D253G/T255D Thermoactinomyces vulgaris
2
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant A243G Thermoactinomyces vulgaris
4.3
-
 2,4-dinitrophenyl-alanyl-alanyl-arginine mutant D253G/T255D Thermoactinomyces vulgaris
6.7
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant A243G Thermoactinomyces vulgaris
6.8
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant G207S/A243G/D253G/T255D Thermoactinomyces vulgaris
11.1
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant A243G/D253G/T255D Thermoactinomyces vulgaris
22.8
-
 benzyloxycarbonyl-alanyl-alanyl-leucine mutant G207S/A243G/T250A/D253G/T255D Thermoactinomyces vulgaris