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 (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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Thermoactinomyces vulgaris | P29068 | - |
- |
Purification (Comment) | Organism |
---|---|
native CpT isolated from inclusion bodies, purified by molecular exclusion chromatography and on cation-exchange resin | Thermoactinomyces vulgaris |
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 | Comment | Organism |
---|---|---|
CPT | - |
Thermoactinomyces vulgaris |
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 |