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Literature summary for 2.5.1.2 extracted from

  • Sikowitz, M.D.; Shome, B.; Zhang, Y.; Begley, T.P.; Ealick, S.E.
    Structure of a Clostridium botulinum C143S thiaminase I/thiamin complex reveals active site architecture (2013), Biochemistry, 52, 7830-7839.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.5.1.2

Crystallization (Commentary)

Crystallization (Comment) Organism
inactive C143S mutant with bound thiamine at 2.2 A resolution. Each monomer contains one thiamine molecule in a V-shaped cleft between the two domains. Tyrosine residues, Tyr46, Tyr48, Tyr80, Tyr252, Tyr269, and Tyr300, form an outer collar of the cleft with most of the active site residues located at the bottom of the cleft. The catalytic cysteine residue Cys143 is located on beta6 Clostridium botulinum

Protein Variants

Protein Variants Comment Organism
C143S inactive Clostridium botulinum
D302N about 20fold decrease in kcat/Km value Clostridium botulinum
D94N about 10000fold decrease in kcat/Km value Clostridium botulinum
E271Q about 3000fold decrease in kcat/Km value Clostridium botulinum
Y46F about 2fold decrease in kcat/Km value Clostridium botulinum
Y80F about 1000fold decrease in kcat/Km value Clostridium botulinum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
0.075
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
0.25
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
0.45
-
 thiamine wild-type, pH 8.0, 37°C Clostridium botulinum
2.1
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
3.5
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum

Organism

Organism UniProt Comment Textmining
Clostridium botulinum
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
thiamine + 2-mercaptoethanol
-
 Clostridium botulinum ?
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
0.23
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum
1.1
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
8.1
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
60.9
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
231
-
 thiamine wild-type, pH 8.0, 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
0.07
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum
0.15
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
0.5
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
26.4
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
252
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
510
-
 thiamine wild-type, pH 8.0, 37°C Clostridium botulinum