Cloned (Comment) | Organism |
---|---|
gene murE, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3):pLysS from plasmid pET2160:murESa, coexpression with chaperone from pREP4groESL | Staphylococcus aureus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme in complex with UDP-MurNAc-Ala-Glu-Lys or with UDP-MurNAc-Ala-Glu-Lys and ADP, from 0.1 M Na-HEPES/MOPS, pH 7.5, 0.09 M nitrate phosphate sulfate mix, 30% w/v PEG550MME/PEG20K, method evaluation, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution, molecular replacement | Staphylococcus aureus |
Protein Variants | Comment | Organism |
---|---|---|
A409R | site-directed mutagenesis, the lytic phenotype observed upon overexpression of the wild-type murESa gene is replicated with the mutant | Staphylococcus aureus |
D406A | site-directed mutagenesis, the mutant shows very low or no activity in vivo | Staphylococcus aureus |
E460A | site-directed mutagenesis, the mutant shows very low or no activity in vivo | Staphylococcus aureus |
N407A | site-directed mutagenesis, inactive mutant | Staphylococcus aureus |
N407R | site-directed mutagenesis, the growth curve of N407R mutant is similar to that of wild-type murESa | Staphylococcus aureus |
P408A | site-directed mutagenesis, the lytic phenotype observed upon overexpression of the wild-type murESa gene is replicated with the mutant | Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.55 | - |
L-lysine | pH 8.6, 37°C, recombinant wild-type enzyme | Staphylococcus aureus | |
2.8 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant D406A | Staphylococcus aureus | |
3.1 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant P408A | Staphylococcus aureus | |
20 | - |
L-lysine | above, pH 8.6, 37°C, recombinant mutant A409R | Staphylococcus aureus | |
20 | - |
L-lysine | above, pH.6, 37°C, recombinant mutant E460A | Staphylococcus aureus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine | Staphylococcus aureus | the enzyme of the Gram-positive bacterium discriminates between L-lysine and D,L-diaminopimelic acid, the predominant amino acid that replaces L-lysine in Gram-negative peptidoglycan. Despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations | ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine | - |
? | |
additional information | Staphylococcus aureus | the third amino acid added to the peptide stem of peptidoglycan is usually either meso-diaminopimelic acid (in most Gram-negative bacteria), EC 6.3.2.13, or L-lysine (in most Gram-positive bacteria), EC 6.3.2.7 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | Q2FZP6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3):pLysS by nickel affinity chromatography, gel filtration, and ion exchange chromatography | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine | the enzyme of the Gram-positive bacterium discriminates between L-lysine and D,L-diaminopimelic acid, the predominant amino acid that replaces L-lysine in Gram-negative peptidoglycan. Despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations | Staphylococcus aureus | ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine | - |
? | |
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine | despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations, substrate discrimination mechanism, overview. Glu460 is crucial for interaction with the epsilon-amino group of the L-lysine substrate | Staphylococcus aureus | ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine | - |
? | |
additional information | the third amino acid added to the peptide stem of peptidoglycan is usually either meso-diaminopimelic acid (in most Gram-negative bacteria), EC 6.3.2.13, or L-lysine (in most Gram-positive bacteria), EC 6.3.2.7 | Staphylococcus aureus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MurE | - |
Staphylococcus aureus |
UDP-N-acetylmuramoyl-L-alanyl-D-glutamate:meso-2,6-diaminopimelate ligase | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Staphylococcus aureus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant D406A | Staphylococcus aureus | |
0.283 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant P408A | Staphylococcus aureus | |
4.833 | - |
L-lysine | pH 8.6, 37°C, recombinant wild-type enzyme | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.6 | - |
assay at | Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Staphylococcus aureus |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in peptidoglycan biosynthesis and cell wall assembly | Staphylococcus aureus |
additional information | ATP-binding site structure and homology, overview. Asn407 makes a series of structurally important hydrogen bond interactions within the context of the architecture of the active site region | Staphylococcus aureus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant D406A | Staphylococcus aureus | |
0.003 | - |
L-lysine | above, pH 8.6, 37°C, recombinant mutant A409R | Staphylococcus aureus | |
0.029 | - |
L-lysine | above, pH.6, 37°C, recombinant mutant E460A | Staphylococcus aureus | |
0.091 | - |
L-lysine | pH 8.6, 37°C, recombinant mutant P408A | Staphylococcus aureus | |
8.787 | - |
L-lysine | pH 8.6, 37°C, recombinant wild-type enzyme | Staphylococcus aureus |