Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | in contrast to Escherichia coli no activation by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Staphylococcus aureus | |
additional information | in contrast to Escherichia coli no activation by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Enterococcus faecalis | |
additional information | in contrast to Escherichia coli no activation by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Enterococcus faecium | |
additional information | in contrast to Escherichia coli no activation by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Helicobacter pylori | |
UDP-MurNAc-Ala | strong activation by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Escherichia coli |
Application | Comment | Organism |
---|---|---|
drug development | glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and is therefore considered as a target for antibacterial drug discovery | Staphylococcus aureus |
drug development | glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and is therefore considered as a target for antibacterial drug discovery | Escherichia coli |
drug development | glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and is therefore considered as a target for antibacterial drug discovery | Enterococcus faecalis |
drug development | glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and is therefore considered as a target for antibacterial drug discovery | Enterococcus faecium |
drug development | glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and is therefore considered as a target for antibacterial drug discovery | Helicobacter pylori |
Cloned (Comment) | Organism |
---|---|
full-length MurI proteins are prepared by expression either as native (Helicobacter pylori, Escherichia coli) or N-terminal 6xHis-tagged (Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium) recombinant proteins in Escherichia coli strains co-expressing the chaperone proteins GroEL/GroES27 and purified by standard chromatographic methods | Staphylococcus aureus |
full-length MurI proteins are prepared by expression either as native (Helicobacter pylori, Escherichia coli) or N-terminal 6xHis-tagged (Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium) recombinant proteins in Escherichia coli strains co-expressing the chaperone proteins GroEL/GroES27 and purified by standard chromatographic methods | Escherichia coli |
full-length MurI proteins are prepared by expression either as native (Helicobacter pylori, Escherichia coli) or N-terminal 6xHis-tagged (Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium) recombinant proteins in Escherichia coli strains co-expressing the chaperone proteins GroEL/GroES27 and purified by standard chromatographic methods | Enterococcus faecalis |
full-length MurI proteins are prepared by expression either as native (Helicobacter pylori, Escherichia coli) or N-terminal 6xHis-tagged (Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium) recombinant proteins in Escherichia coli strains co-expressing the chaperone proteins GroEL/GroES27 and purified by standard chromatographic methods | Enterococcus faecium |
full-length MurI proteins are prepared by expression either as native (Helicobacter pylori, Escherichia coli) or N-terminal 6xHis-tagged (Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium) recombinant proteins in Escherichia coli strains co-expressing the chaperone proteins GroEL/GroES27 and purified by standard chromatographic methods | Helicobacter pylori |
Crystallization (Comment) | Organism |
---|---|
crystal structures of MurI of Helicobacter pylori, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are analysed under similar, physiologically relevant conditons. MurI of Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium all form homodimeric structures. In all these structures, monomers oligomerize in a tail-to-tail orientation with active sites opposed and fully exposed to solvent | Staphylococcus aureus |
crystal structures of MurI of Helicobacter pylori, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are analysed under similar, physiologically relevant conditons. MurI of Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium all form homodimeric structures. In all these structures, monomers oligomerize in a tail-to-tail orientation with active sites opposed and fully exposed to solvent | Enterococcus faecalis |
crystal structures of MurI of Helicobacter pylori, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are analysed under similar, physiologically relevant conditons. MurI of Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium all form homodimeric structures. In all these structures, monomers oligomerize in a tail-to-tail orientation with active sites opposed and fully exposed to solvent | Enterococcus faecium |
crystal structures of MurI of Helicobacter pylori, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are analysed under similar, physiologically relevant conditons. The Escherichia coli MurI co-crystallizes as a monomer with both L-glutamate and its activator UDP-MurNAc-Ala. The activator binds in the hinge region on the side opposite to the catalytically active site through contacts between the uracil ring system and domain B and through specific salt bridge interactions with R104 in domain A and the alanyl moiety of the activator, consistent with the strict requirement of the alanine and uracil moieties for activation | Escherichia coli |
the different kinetic profiles of MurI enzymes across the species suggest fundamental structural differences and therefore, crystal structures of MurI of Helicobacter pylori, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium are analysed under similar, physiologically relevant conditons. The Helicobacter pylori MurI enzyme also forms a homodimer but with the active sites in close proximity in a face-to-face orientation | Helicobacter pylori |
Protein Variants | Comment | Organism |
---|---|---|
A75T | Ki: 0.661 mM (inhibitor: D-glutamate). Turnover number: 1.78/sec (substrate: L-glutamate), 0.065/sec (substrate: D-glutamate). Km: 7.4 mM (substrate: L-glutamate), Km: 0.275 mM (substrate: D-glutamate) | Helicobacter pylori |
E151T | Ki: 100 mM (value above 100 for inhibitor: D-glutamate). Turnover number: 0.08/sec (substrate: D-glutamate), 2.26/sec (substrate: L-glutamate). Km: 7.36 mM (substrate: L-glutamate), Km: 0.282 mM (substrate: D-glutamate) | Helicobacter pylori |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5-methyl-7-(2-methylpropyl)-2-(naphthalen-1-ylmethyl)-3-pyridin-4-yl-2H-pyrazolo[3,4-d]pyrimidine-4,6(5H,7H)-dione | a pyrazolopyrimidinedione analogue identified by a high-throughput screen demonstrates inhibition with excellent selectivity for MurI of Helicobacter pylori, it is time-independent, fully-reversible and insensitive to changes in enzyme or detergent concentration | Helicobacter pylori | |
D-glutamate | MurI of Helicobacter pylori is strongly inhibited by D-glutamate | Helicobacter pylori | |
additional information | in contrast to MurI of Helicobacter pylori no inhibition by D-glutamate | Enterococcus faecalis | |
additional information | in contrast to MurI of Helicobacter pylori no inhibition by D-glutamate | Enterococcus faecium | |
additional information | in contrast to MurI of Helicobacter pylori no inhibition by D-glutamate | Escherichia coli | |
additional information | in contrast to MurI of Helicobacter pylori no inhibition by D-glutamate | Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
D-glutamate | - |
Staphylococcus aureus | |
0.063 | - |
D-glutamate | - |
Helicobacter pylori | |
0.24 | - |
D-glutamate | - |
Enterococcus faecium | |
0.25 | - |
D-glutamate | - |
Enterococcus faecalis | |
0.275 | - |
D-glutamate | deletion mutant A75T | Helicobacter pylori | |
0.282 | - |
D-glutamate | deletion mutant E151T | Helicobacter pylori | |
0.74 | - |
L-glutamate | demonstrates a high degree of asymmetry in substrate processing with the Michaelis constant for D-glutamate approximately tenfold lower than for L-glutamate | Helicobacter pylori | |
1.1 | - |
L-glutamate | - |
Enterococcus faecium | |
1.2 | - |
L-glutamate | - |
Escherichia coli | |
1.2 | - |
L-glutamate | - |
Enterococcus faecalis | |
2.1 | - |
D-glutamate | - |
Escherichia coli | |
4.6 | - |
L-glutamate | - |
Staphylococcus aureus | |
7.36 | - |
L-glutamate | deletion mutant E151T | Helicobacter pylori | |
7.4 | - |
L-glutamate | deletion mutant A75T | Helicobacter pylori |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Enterococcus faecalis | - |
- |
- |
Enterococcus faecium | - |
- |
- |
Escherichia coli | - |
- |
- |
Helicobacter pylori | - |
- |
- |
Staphylococcus aureus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
enzyme is purified by using standard chromatographic methods | Staphylococcus aureus |
enzyme is purified by using standard chromatographic methods | Escherichia coli |
enzyme is purified by using standard chromatographic methods | Enterococcus faecalis |
enzyme is purified by using standard chromatographic methods | Enterococcus faecium |
enzyme is purified by using standard chromatographic methods | Helicobacter pylori |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
Escherichia coli MurI is monomeric in solution and shows extremely low intrinsic activity in either direction (symmetrical kinetic profile), but catalytic turnover is upregulated over 1000fold by UDP-MurNAc-Ala, the product of the preceding enzyme in the peptidoglycan biosynthetic pathway | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | - |
Escherichia coli | D-glutamate | - |
r | |
L-glutamate | - |
Helicobacter pylori | D-glutamate | - |
r | |
L-glutamate | MurI enzymes from the Gram-positive species Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium share similar biophysical and biochemical characteristics that are distinct from Escherichia coli and Helicobacter pylori MurI | Staphylococcus aureus | D-glutamate | - |
r | |
L-glutamate | MurI enzymes from the Gram-positive species Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium share similar biophysical and biochemical characteristics that are distinct from Escherichia coli and Helicobacter pylori MurI | Enterococcus faecalis | D-glutamate | - |
r | |
L-glutamate | MurI enzymes from the Gram-positive species Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium share similar biophysical and biochemical characteristics that are distinct from Escherichia coli and Helicobacter pylori MurI | Enterococcus faecium | D-glutamate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | method: gel filtration. The Helicobacter pylori MurI enzyme also forms a homodimer but with the active sites in close proximity in a face-to-face orientation | Helicobacter pylori |
monomer | gel filtration | Staphylococcus aureus |
monomer | gel filtration | Escherichia coli |
monomer | gel filtration | Enterococcus faecalis |
monomer | gel filtration | Enterococcus faecium |
Synonyms | Comment | Organism |
---|---|---|
glutamate racemase | - |
Staphylococcus aureus |
glutamate racemase | - |
Escherichia coli |
glutamate racemase | - |
Enterococcus faecalis |
glutamate racemase | - |
Enterococcus faecium |
glutamate racemase | - |
Helicobacter pylori |
MurI | - |
Staphylococcus aureus |
MurI | - |
Escherichia coli |
MurI | - |
Enterococcus faecalis |
MurI | - |
Enterococcus faecium |
MurI | - |
Helicobacter pylori |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.065 | - |
D-glutamate | deletion mutant A75T | Helicobacter pylori | |
0.08 | - |
D-glutamate | deletion mutant E151T | Helicobacter pylori | |
0.2 | - |
D-glutamate | - |
Helicobacter pylori | |
0.41 | - |
L-glutamate | - |
Escherichia coli | |
0.41 | - |
D-glutamate | - |
Enterococcus faecalis | |
0.56 | - |
D-glutamate | - |
Staphylococcus aureus | |
1.05 | - |
L-glutamate | - |
Helicobacter pylori | |
1.78 | - |
L-glutamate | deletion mutant A75T | Helicobacter pylori | |
2.26 | - |
L-glutamate | deletion mutant E151T | Helicobacter pylori | |
3.4 | - |
L-glutamate | - |
Enterococcus faecium | |
8.5 | - |
L-glutamate | - |
Staphylococcus aureus | |
11.73 | - |
D-glutamate | - |
Enterococcus faecalis | |
12.16 | - |
L-glutamate | - |
Escherichia coli | |
15 | - |
D-glutamate | - |
Enterococcus faecium | |
25 | - |
L-glutamate | - |
Enterococcus faecalis | |
36.66 | - |
L-glutamate | - |
Enterococcus faecium | |
43.33 | - |
D-glutamate | - |
Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0058 | - |
D-glutamate | - |
Helicobacter pylori | |
0.661 | - |
D-glutamate | deletion mutant A75T | Helicobacter pylori | |
100 | - |
D-glutamate | value above 100 for deletion mutant E151K | Helicobacter pylori |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0014 | - |
a pyrazolopyrimidinedione analogue (compound A) identified by a high-throughput screen demonstrates inhibition with excellent selectivity for MurI of Helicobacter pylori, it is time-independent, fully-reversible and insensitive to changes in enzyme or detergent concentration | Helicobacter pylori | pyrazolopyrimidinedione analogue | |
0.4 | - |
value above 0.4, a pyrazolopyrimidinedione analogue (compound A) identified by a high-throughput screen demonstrates inhibition with excellent selectivity for MurI of Helicobacter pylori but not for MurI of Escherichia coli | Escherichia coli | pyrazolopyrimidinedione analogue | |
0.4 | - |
value above 0.4, a pyrazolopyrimidinedione analogue (compound A) identified by a high-throughput screen demonstrates inhibition with excellent selectivity for MurI of Helicobacter pylori but not for MurI of Staphylococcus aureus | Staphylococcus aureus | pyrazolopyrimidinedione analogue |