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Literature summary for 3.4.24.3 extracted from

  • Eckhard, U.; Schoenauer, E.; Ducka, P.; Briza, P.; Nuess, D.; Brandstetter, H.
    Biochemical characterization of the catalytic domains of three different clostridial collagenases (2009), Biol. Chem., 390, 11-18.
    View publication on PubMed
Search for more literature for 3.4.24.3

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Clostridium tetani
expression in Escherichia coli Hathewaya histolytica

Protein Variants

Protein Variants Comment Organism
G426V isoform ColH, complete loss of enzymatic activity Hathewaya histolytica
additional information addition of an N-terminal His-tag. The Michaelis-Menten constant of ColG and ColH increases by approximately 50% and 60%, respectively Hathewaya histolytica
additional information addition of an N-terminal His-tag. The Michaelis-Menten constant of ColT increases by approximately 60%. Expression of a truncated construct Y53-A506 which stops shortly after the third zinc binding residue and consitstent with the prediction of a minimal catalytic domain. The construct shows no catalytic activity at all, despite containing all zinc binding residues including the catalytic glutamate that is supposed to act as general base Clostridium tetani

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline The half maximal inhibitory concentrations of the catalytic domains for 1,10-phenanthroline are significantly decreased in the presence of the N-terminal His6-tag Clostridium tetani
1,10-phenanthroline The half maximal inhibitory concentrations of the catalytic domains for 1,10-phenanthroline are significantly decreased for both isoforms ColG and ColH in the presence of the N-terminal His6-tag Hathewaya histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.269
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColH, pH 7.5, 25°C Hathewaya histolytica
0.425
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColH with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica
0.84
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColG, pH 7.5, 25°C Hathewaya histolytica
0.97
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColT, pH 7.5, 25°C Clostridium tetani
1.24
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColT with N-terminal His6-tag, pH 7.5, 25°C Clostridium tetani
1.26
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColG with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica

Metals/Ions

Metals/Ions Comment Organism Structure
Zinc expression of a truncated construct Y53-A506 which stops shortly after the third zinc binding residue and consitstent with the prediction of a minimal catalytic domain. The construct shows no catalytic activity at all, despite containing all zinc binding residues including the catalytic glutamate that is supposed to act as general base Clostridium tetani

Organism

Organism UniProt Comment Textmining
Clostridium tetani
-
-
-
Hathewaya histolytica
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
furylacryloyl-Leu-Gly-Pro-Ala + H2O
-
 Clostridium tetani furylacryloyl-Leu + Gly-Pro-Ala
-
 ?
furylacryloyl-Leu-Gly-Pro-Ala + H2O
-
 Hathewaya histolytica furylacryloyl-Leu + Gly-Pro-Ala
-
 ?

Synonyms

Synonyms Comment Organism
ColG
-
 Hathewaya histolytica
ColH
-
 Hathewaya histolytica
collagenase H
-
 Hathewaya histolytica
collagenase T
-
 Clostridium tetani
ColT
-
 Clostridium tetani

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.092
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColG with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica
0.11
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColG, pH 7.5, 25°C Hathewaya histolytica
3.65
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColT with N-terminal His6-tag, pH 7.5, 25°C Clostridium tetani
7.24
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColT, pH 7.5, 25°C Clostridium tetani
13.6
-
 furylacryloyl-Leu-Gly-Pro-Ala isoform ColH with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica
15.9
-
 furylacryloyl-Leu-Gly-Pro-Ala wild-type ColH, pH 7.5, 25°C Hathewaya histolytica

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.025
-
 isoform ColH with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica 1,10-phenanthroline
0.04
-
 isoform ColT with N-terminal His6-tag, pH 7.5, 25°C Clostridium tetani 1,10-phenanthroline
0.055
-
 wild-type ColH, pH 7.5, 25°C Hathewaya histolytica 1,10-phenanthroline
0.1
-
 isoform ColG with N-terminal His6-tag, pH 7.5, 25°C Hathewaya histolytica 1,10-phenanthroline
0.15
-
 wild-type ColG, pH 7.5, 25°C Hathewaya histolytica 1,10-phenanthroline
0.15
-
 wild-type ColT, pH 7.5, 25°C Clostridium tetani 1,10-phenanthroline