Literature summary extracted from
Samland, A.K.; Amrhein, N.; Macheroux, P.
Lysine 22 in UDP-N-acetylglucosamine enolpyruvyl transferase from Enterobacter cloacae is crucial for enzymatic activity and the formation of covalent adducts with the substrate phosphoenolpyruvate and the antibiotic fosfomycin (1999), Biochemistry, 38, 13162-13169.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
2.5.1.7 |
dithiothreitol |
included in the assay reaction mixture |
Enterobacter cloacae |
|
Application
EC Number |
Application |
Comment |
Organism |
---|
2.5.1.7 |
medicine |
enzyme is a target for the antibiotic fosfomycin |
Enterobacter cloacae |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.5.1.7 |
expression of wild-type and mutants in Escherichia coli |
Enterobacter cloacae |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
2.5.1.7 |
C115S |
- |
Enterobacter cloacae |
2.5.1.7 |
K22E |
site-directed mutagenesis, exchange of conserved Lys residue located near the active site and involved in substrate binding leading to conformational changes, shows less than 0.5% activity compared to the wild-type, altered UDP-GlcNAc binding, highly reduced formation of covalent adduct between active site Cys115 and phosphoenolpyruvate or inhibitor fosfomycin |
Enterobacter cloacae |
2.5.1.7 |
K22R |
site-directed mutagenesis, exchange of conserved Lys residue located near the active site and involved in substrate binding leading to conformational changes, shows less than 0.5% activity compared to the wild-type, slightly reduced formation of covalent adduct between active site Cys115 and phosphoenolpyruvate or inhibitor fosfomycin |
Enterobacter cloacae |
2.5.1.7 |
K22V |
site-directed mutagenesis, exchange of conserved Lys residue located near the active site and involved in substrate binding leading to conformational changes, shows less than 0.5% activity compared to the wild-type, reduced formation of covalent adduct between active site Cys115 and phosphoenolpyruvate or inhibitor fosfomycin |
Enterobacter cloacae |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
2.5.1.7 |
fosfomycin |
- |
Enterobacter cloacae |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.5.1.7 |
Enterobacter cloacae |
P33038 |
MurA |
- |
2.5.1.7 |
Enterobacter cloacae DSM 30054 |
P33038 |
MurA |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.5.1.7 |
recombinant wild-type and mutants from Escherichia coli |
Enterobacter cloacae |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
2.5.1.7 |
phosphoenolpyruvate + UDP-N-acetyl-alpha-D-glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine |
Lys22 is located near the active site and involved in substrate binding |
Enterobacter cloacae |
|
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
2.5.1.7 |
additional information |
- |
activity of K22 mutants compared to wild-type in forward and reverse reaction, coupled assay |
Enterobacter cloacae |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
2.5.1.7 |
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine |
substrate binding structure |
Enterobacter cloacae |
phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine |
- |
r |
|
2.5.1.7 |
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine |
thermodynamic parameters of substrate binding, wild-type, C115S and K22 mutants |
Enterobacter cloacae |
phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine |
- |
r |
|
2.5.1.7 |
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine |
substrate binding structure |
Enterobacter cloacae DSM 30054 |
phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine |
- |
r |
|
2.5.1.7 |
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine |
thermodynamic parameters of substrate binding, wild-type, C115S and K22 mutants |
Enterobacter cloacae DSM 30054 |
phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine |
- |
r |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
2.5.1.7 |
More |
structure |
Enterobacter cloacae |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
2.5.1.7 |
25 |
- |
assay at |
Enterobacter cloacae |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
2.5.1.7 |
7.4 |
- |
assay at |
Enterobacter cloacae |