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Literature summary for 3.4.24.69 extracted from

  • Chen, S.; Wan, H.Y.
    Molecular mechanisms of substrate recognition and specificity of botulinum neurotoxin serotype F (2011), Biochem. J., 433, 277-284.
    View publication on PubMed
Search for more literature for 3.4.24.69

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Clostridium botulinum

Protein Variants

Protein Variants Comment Organism
E200A Km (VAMP-2) similar to wild-type, kcat strongly decreased compared to wild-type Clostridium botulinum
E315A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
I52A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
K172A Km (VAMP-2) increased compared to wild-type, kcat moderately decreased compared to wild-type Clostridium botulinum
L173A Km (VAMP-2) increased compared to wild-type, kcat moderately decreased compared to wild-type Clostridium botulinum
P25A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
R171A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
R240A Km (VAMP-2) similar to wild-type, kcat decreased compared to wild-type Clostridium botulinum
R263A Km (VAMP-2) similar to wild-type, kcat decreased compared to wild-type Clostridium botulinum
S147A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
S224A Km (VAMP-2) similar to wild-type, kcat decreased compared to wild-type Clostridium botulinum
V137A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
W319A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
W44A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
Y113A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
Y133A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
Y168A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
Y26A Km (VAMP-2) increased compared to wild-type, kcat decreased compared to wild-type Clostridium botulinum
Y322A Km (VAMP-2) increased compared to wild-type, kcat similar to wild-type Clostridium botulinum
Y368A Km (VAMP-2) similar to wild-type, kcat strongly decreased compared to wild-type Clostridium botulinum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0067
-
 VAMP-2 pH 7.6, 37°C, mutant R263A Clostridium botulinum
0.0069
-
 VAMP-2 pH 7.6, 37°C, mutant R240A Clostridium botulinum
0.0069
-
 VAMP-2 pH 7.6, 37°C, mutant Y368A Clostridium botulinum
0.007
-
 VAMP-2 pH 7.6, 37°C, wild-type Clostridium botulinum
0.0072
-
 VAMP-2 pH 7.6, 37°C, mutant E200A Clostridium botulinum
0.0075
-
 VAMP-2 pH 7.6, 37°C, mutant S224A Clostridium botulinum
0.0158
-
 VAMP-2 pH 7.6, 37°C, mutant L173A Clostridium botulinum
0.0356
-
 VAMP-2 pH 7.6, 37°C, mutant I52A Clostridium botulinum
0.038
-
 VAMP-2 pH 7.6, 37°C, mutant S147A Clostridium botulinum
0.0556
-
 VAMP-2 pH 7.6, 37°C, mutant Y322A Clostridium botulinum
0.0596
-
 VAMP-2 pH 7.6, 37°C, mutant W319A Clostridium botulinum
0.0702
-
 VAMP-2 pH 7.6, 37°C, mutant Y26A Clostridium botulinum
0.0737
-
 VAMP-2 pH 7.6, 37°C, mutant E315A Clostridium botulinum
0.0753
-
 VAMP-2 pH 7.6, 37°C, mutant K172A Clostridium botulinum
0.0798
-
 VAMP-2 pH 7.6, 37°C, mutant Y113A Clostridium botulinum
0.0838
-
 VAMP-2 pH 7.6, 37°C, mutant R171A Clostridium botulinum
0.0857
-
 VAMP-2 pH 7.6, 37°C, mutant Y168A Clostridium botulinum
0.1308
-
 VAMP-2 pH 7.6, 37°C, mutant Y133A Clostridium botulinum
0.1405
-
 VAMP-2 pH 7.6, 37°C, mutant V137A Clostridium botulinum
0.1804
-
 VAMP-2 pH 7.6, 37°C, mutant P25A Clostridium botulinum
0.218
-
 VAMP-2 pH 7.6, 37°C, mutant W44A Clostridium botulinum

Organism

Organism UniProt Comment Textmining
Clostridium botulinum
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
VAMP-2 + H2O initial substrate recognition is mediated through sequential binding of VAMP-2 to the B1, B2 and B3 pockets in LC/F (light chain of BoNT serotype F), which directed VAMP-2 to the active site of LC/F and stabilized the active site substrate recognition, where the P2, P1' and P2' sites of VAMP-2 are specifically recognized by the S2, S1' and S2' pockets of LC/F to promote substrate hydrolysis Clostridium botulinum ?
-
 ?

Synonyms

Synonyms Comment Organism
BoNT serotype F
-
 Clostridium botulinum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Clostridium botulinum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0014
-
 VAMP-2 pH 7.6, 37°C, mutant E200A Clostridium botulinum
0.048
-
 VAMP-2 pH 7.6, 37°C, mutant Y368A Clostridium botulinum
0.23
-
 VAMP-2 pH 7.6, 37°C, mutant Y26A Clostridium botulinum
0.52
-
 VAMP-2 pH 7.6, 37°C, mutant R171A Clostridium botulinum
0.52
-
 VAMP-2 pH 7.6, 37°C, mutant R240A Clostridium botulinum
0.58
-
 VAMP-2 pH 7.6, 37°C, mutant Y168A Clostridium botulinum
1.1
-
 VAMP-2 pH 7.6, 37°C, mutant R263A Clostridium botulinum
2.3
-
 VAMP-2 pH 7.6, 37°C, mutant P25A Clostridium botulinum
3.2
-
 VAMP-2 pH 7.6, 37°C, mutant I52A Clostridium botulinum
4.2
-
 VAMP-2 pH 7.6, 37°C, mutant S224A Clostridium botulinum
4.5
-
 VAMP-2 pH 7.6, 37°C, mutant V137A Clostridium botulinum
5.2
-
 VAMP-2 pH 7.6, 37°C, mutant Y133A Clostridium botulinum
40.8
-
 VAMP-2 pH 7.6, 37°C, mutant L173A Clostridium botulinum
120.5
-
 VAMP-2 pH 7.6, 37°C, mutant K172A Clostridium botulinum
198.8
-
 VAMP-2 pH 7.6, 37°C, mutant S147A Clostridium botulinum
200.7
-
 VAMP-2 pH 7.6, 37°C, mutant W319A Clostridium botulinum
203.5
-
 VAMP-2 pH 7.6, 37°C, mutant W44A Clostridium botulinum
220.8
-
 VAMP-2 pH 7.6, 37°C, wild-type Clostridium botulinum
225.8
-
 VAMP-2 pH 7.6, 37°C, mutant E315A Clostridium botulinum
240.8
-
 VAMP-2 pH 7.6, 37°C, mutant Y113A Clostridium botulinum
240.8
-
 VAMP-2 pH 7.6, 37°C, mutant Y322A Clostridium botulinum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
 assay at Clostridium botulinum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.19
-
 VAMP-2 pH 7.6, 37°C, mutant E200A Clostridium botulinum
0.32
-
 VAMP-2 pH 7.6, 37°C, mutant Y26A Clostridium botulinum
6.2
-
 VAMP-2 pH 7.6, 37°C, mutant R171A Clostridium botulinum
6.9
-
 VAMP-2 pH 7.6, 37°C, mutant Y168A Clostridium botulinum
7
-
 VAMP-2 pH 7.6, 37°C, mutant Y368A Clostridium botulinum
13
-
 VAMP-2 pH 7.6, 37°C, mutant P25A Clostridium botulinum
19
-
 VAMP-2 pH 7.6, 37°C, mutant V137A Clostridium botulinum
32
-
 VAMP-2 pH 7.6, 37°C, mutant Y133A Clostridium botulinum
75
-
 VAMP-2 pH 7.6, 37°C, mutant R240A Clostridium botulinum
89
-
 VAMP-2 pH 7.6, 37°C, mutant I52A Clostridium botulinum
160
-
 VAMP-2 pH 7.6, 37°C, mutant R263A Clostridium botulinum
560
-
 VAMP-2 pH 7.6, 37°C, mutant S224A Clostridium botulinum
920
-
 VAMP-2 pH 7.6, 37°C, mutant W44A Clostridium botulinum
1600
-
 VAMP-2 pH 7.6, 37°C, mutant K172A Clostridium botulinum
2700
-
 VAMP-2 pH 7.6, 37°C, mutant L173A Clostridium botulinum
3000
-
 VAMP-2 pH 7.6, 37°C, mutant Y113A Clostridium botulinum
3100
-
 VAMP-2 pH 7.6, 37°C, mutant E315A Clostridium botulinum
3400
-
 VAMP-2 pH 7.6, 37°C, mutant W319A Clostridium botulinum
4330
-
 VAMP-2 pH 7.6, 37°C, mutant Y322A Clostridium botulinum
4800
-
 VAMP-2 pH 7.6, 37°C, mutant S147A Clostridium botulinum
31100
-
 VAMP-2 pH 7.6, 37°C, wild-type Clostridium botulinum