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Literature summary for 3.4.24.69 extracted from
- Unique substrate recognition mechanism of the botulinum neurotoxin D light chain (2013), J. Biol. Chem., 288, 27881-27887.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3)-RIL cells | Clostridium botulinum |
Protein Variants
| 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 [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/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| vesicle-associated membrane protein-2 + H2O | Clostridium botulinum | - |
? | - |
? |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Clostridium botulinum | P19321 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| vesicle-associated membrane protein-2 + H2O | - |
Clostridium botulinum | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BoNT/D | - |
Clostridium botulinum |
| botulinum neurotoxin D light chain | - |
Clostridium botulinum |
| LC/D | - |
Clostridium botulinum |
Turnover Number [1/s]
| 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 [mM/s]
| 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 |
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