Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 cells | Trypanosoma brucei |
Protein Variants | Comment | Organism |
---|---|---|
E607Q | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E607Q/E609Q | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609A | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609F | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609L | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609Q | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609Q/E610Q | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E609R | the mutant shows reduced kcat/Km compared to the wild type enzyme | Trypanosoma brucei |
E610Q | the mutant shows reduced kcat/Km compared to the wild type enzyme, the mutation results in a marked decrease in thermal stability, the mutant is inactivated to 63% of the control by 3 M urea | Trypanosoma brucei |
General Stability | Organism |
---|---|
the wild type enzyme is not digested by trypsin at 25°C | Trypanosoma brucei |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | reducing reagent such as dithiothreitol has no effect on the inactivation of wild type and mutant OPBs by heat and urea treatment | Trypanosoma brucei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00268 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei | |
0.00334 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei | |
0.00525 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
0.00528 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei | |
0.00568 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
0.00586 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
0.00637 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
0.0069 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
0.00728 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
0.00807 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei | |
0.00904 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
0.00913 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
0.0101 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
0.0113 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
0.0127 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
0.0127 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
0.015 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
0.0173 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
0.0184 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei | |
0.0215 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
84000 | - |
2 * 84000, calculated from amino acid sequence | Trypanosoma brucei |
160000 | - |
gel filtration | Trypanosoma brucei |
Organic Solvent | Comment | Organism |
---|---|---|
urea | the wild type enzyme is resistant to treatment with 3 M urea | Trypanosoma brucei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography | Trypanosoma brucei |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0001 | - |
mutant enzyme E607Q/E609Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
0.000323 | - |
mutant enzyme E607Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
0.011 | - |
mutant enzyme E609Q/E610Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
0.0209 | - |
mutant enzyme E610Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
0.0221 | - |
mutant enzyme E609Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
0.0279 | - |
wild type enzyme, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C | Trypanosoma brucei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
adrenocorticotropic hormone + H2O | - |
Trypanosoma brucei | SYSMEHFR + WGK + WGKPVGK + VYP | - |
? | |
benzoyl-L-Arg-7-amido-4-methylcoumarin + H2O | - |
Trypanosoma brucei | benzoyl-L-Arg + 7-amino-4-methylcoumarin | - |
? | |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin + H2O | - |
Trypanosoma brucei | benzyloxycarbonyl-L-Arg + 7-amino-4-methylcoumarin | - |
? | |
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O | - |
Trypanosoma brucei | benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin | - |
? | |
Glucagon + H2O | wild type enzyme cleaves Lys12-Tyr13, Arg17-Arg18, and Arg18-Ala19 bonds of glucagon | Trypanosoma brucei | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 84000, calculated from amino acid sequence | Trypanosoma brucei |
Synonyms | Comment | Organism |
---|---|---|
oligopeptidase B | - |
Trypanosoma brucei |
OPB | - |
Trypanosoma brucei |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 53 | following treatment at 45°C for 20 min the relative activity of wild type enzyme is 89%, the melting temperature of the wild type enzyme is at 53.3°C | Trypanosoma brucei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.758 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
1.24 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
1.42 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei | |
1.46 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
1.58 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
1.64 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
1.77 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
4.98 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
5.03 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
5.37 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei | |
5.42 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
6.84 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
7.81 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei | |
8.29 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei | |
9.36 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
9.53 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
11.2 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
13.3 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei | |
15.2 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
15.8 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
50 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
67 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
77 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei | |
83 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q, at 37°C | Trypanosoma brucei | |
129 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
217 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E607Q/E609Q, at 37°C | Trypanosoma brucei | |
250 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609R, at 37°C | Trypanosoma brucei | |
265 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609F, at 37°C | Trypanosoma brucei | |
551 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
647 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
677 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
750 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q/E610Q, at 37°C | Trypanosoma brucei | |
790 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609L, at 37°C | Trypanosoma brucei | |
1030 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609A, at 37°C | Trypanosoma brucei | |
1480 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei | |
1650 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
1960 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E609Q, at 37°C | Trypanosoma brucei | |
2480 | - |
benzoyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei | |
2590 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | mutant enzyme E610Q, at 37°C | Trypanosoma brucei | |
4960 | - |
benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin | wild type enzyme, at 37°C | Trypanosoma brucei |