Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)-RIL cells | Clostridium botulinum |
Protein Variants | Comment | Organism |
---|---|---|
F50A | the mutant shows wild type activity | Clostridium botulinum |
F50A/I191A | the mutant shows 60fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
F50D | the mutant shows 4fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
F50D/I191D | the mutant shows 400fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
H132A | the mutant shows wild type activity | Clostridium botulinum |
H132D | inactive | Clostridium botulinum |
H132Q | the mutant shows 100fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
I151A | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
I151D | the mutant shows 1000fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
I152D | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
I191A | the mutant shows wild type activity | Clostridium botulinum |
I191D | the mutant shows 4fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
L200A | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
L200D | the mutant shows 8fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
P154D | the mutant shows 4fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
R23A | the mutant shows wild type activity | Clostridium botulinum |
R23D/H132A | the mutant shows 25fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
R372A | the mutant shows 40fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
R63A | the mutant shows 10fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
R63E | the mutant shows 50fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
V148/I151A | the mutant shows 15fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
V148A | the mutant shows wild type activity | Clostridium botulinum |
V148D | the mutant shows wild type activity | Clostridium botulinum |
W315A | the mutant shows 20fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
W315D | the mutant shows 40fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
W44A/I152A/P154A | the mutant shows 20fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
W44D | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
Y168A | the mutant shows wild type activity | Clostridium botulinum |
Y168A/L200A | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
Y168D | the mutant shows 2fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
Y168D/L200D | the mutant shows 60fold activity reduction compared to the wild type enzyme | Clostridium botulinum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00209 | - |
vesicle-associated membrane protein-2 | mutant enzyme R372A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00219 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00226 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63E, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.0024 | - |
vesicle-associated membrane protein-2 | mutant enzyme L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00291 | - |
vesicle-associated membrane protein-2 | wild type enzyme, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00324 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.0033 | - |
vesicle-associated membrane protein-2 | mutant enzyme Y168D/L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00356 | - |
vesicle-associated membrane protein-2 | mutant enzyme I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00446 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D/I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00481 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50A/I191A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.00796 | - |
vesicle-associated membrane protein-2 | mutant enzyme I151D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.0099 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.01186 | - |
vesicle-associated membrane protein-2 | mutant enzyme P154D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.03245 | - |
vesicle-associated membrane protein-2 | mutant enzyme W44A/I152A/P154A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.03287 | - |
vesicle-associated membrane protein-2 | mutant enzyme R23D/H132A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.03556 | - |
vesicle-associated membrane protein-2 | mutant enzyme H132Q, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.04763 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
vesicle-associated membrane protein-2 + H2O | Clostridium botulinum | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium botulinum | P19321 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
vesicle-associated membrane protein-2 + H2O | - |
Clostridium botulinum | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BoNT/D | - |
Clostridium botulinum |
botulinum neurotoxin D light chain | - |
Clostridium botulinum |
LC/D | - |
Clostridium botulinum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
vesicle-associated membrane protein-2 | mutant enzyme I151D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.04 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D/I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.12 | - |
vesicle-associated membrane protein-2 | mutant enzyme Y168D/L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.15 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63E, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.17 | - |
vesicle-associated membrane protein-2 | mutant enzyme R372A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.28 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50A/I191A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.4 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.81 | - |
vesicle-associated membrane protein-2 | mutant enzyme L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
0.86 | - |
vesicle-associated membrane protein-2 | mutant enzyme H132Q, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
1.4 | - |
vesicle-associated membrane protein-2 | mutant enzyme I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
1.52 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
1.6 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
2.38 | - |
vesicle-associated membrane protein-2 | mutant enzyme R23D/H132A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
3.92 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
5.86 | - |
vesicle-associated membrane protein-2 | mutant enzyme W44A/I152A/P154A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
6.35 | - |
vesicle-associated membrane protein-2 | mutant enzyme P154D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
6.88 | - |
vesicle-associated membrane protein-2 | wild type enzyme, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.5 | - |
vesicle-associated membrane protein-2 | mutant enzyme I151D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
8.9 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D/I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
24 | - |
vesicle-associated membrane protein-2 | mutant enzyme H132Q, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
36 | - |
vesicle-associated membrane protein-2 | mutant enzyme Y168D/L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
58 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50A/I191A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
66 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63E, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
72 | - |
vesicle-associated membrane protein-2 | mutant enzyme R23D/H132A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
81 | - |
vesicle-associated membrane protein-2 | mutant enzyme R372A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
82 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
150 | - |
vesicle-associated membrane protein-2 | mutant enzyme W315A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
180 | - |
vesicle-associated membrane protein-2 | mutant enzyme R63A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
180 | - |
vesicle-associated membrane protein-2 | mutant enzyme W44A/I152A/P154A, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
340 | - |
vesicle-associated membrane protein-2 | mutant enzyme L200D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
390 | - |
vesicle-associated membrane protein-2 | mutant enzyme I191D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
490 | - |
vesicle-associated membrane protein-2 | mutant enzyme F50D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
540 | - |
vesicle-associated membrane protein-2 | mutant enzyme P154D, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum | |
2360 | - |
vesicle-associated membrane protein-2 | wild type enzyme, in 10 mM Tris-HCl (pH 7.6) and 20 mM NaCl, at 37°C | Clostridium botulinum |