Any feedback?
Literature summary extracted from
- Exploring the structure of glutamate racemase from Mycobacterium tuberculosis as a template for anti-mycobacterial drug discovery (2016), Biochem. J., 473, 1267-1280 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.1.1.3 | gene murI, recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Mycolicibacterium smegmatis |
| 5.1.1.3 | gene murI, recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) pLysS | Mycobacterium tuberculosis |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 5.1.1.3 | purified recombinant enzyme complexed with D-glutamate, hanging drop vapour diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 20 mM Tris HCl, pH 8.0, and 1 mM D-glutamate, with 0.002 ml of 18% PEG3350, 0.2 M NaI, 4°C, 4 days, X-ray diffraction structure determination and analysis at 1.76 A resolution | Mycolicibacterium smegmatis |
| 5.1.1.3 | purified recombinant enzyme complexed with D-glutamate, microbatch-underoil method, mixing of 0.002 ml of 5 mg/ml protein in 20 mM Tris HCl, pH 8.0, and 1 mM D-glutamate, with 0.001 ml of crystallizing solution containing 0.1 M HEPES, pH 7.5, and 20% PEG 10000, 18°C, 14 days, X-ray diffraction structure determination and analysis at 2.30 A resolution | Mycobacterium tuberculosis |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.1.1.3 | D26R/R105A/G194E | site-directed mutagenesis, mutation of the interface affording a soluble and active enzyme | Mycobacterium tuberculosis |
| 5.1.1.3 | D26R/R105A/G194R | site-directed mutagenesis, mutation of the interface affording a soluble and active enzyme | Mycobacterium tuberculosis |
| 5.1.1.3 | additional information | site-directed mutagenesis of the enzyme's interface affording a soluble and active enzyme | Mycolicibacterium smegmatis |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.1.1.3 | additional information | inhibitor screening, compounds designed to target other glutamate racemases have been screened but do not inhibit mycobacterial MurI, suggesting that a different drug design effort will be needed to develop inhibitors. A distinct type of MurI dimer arrangement is observed in both structures, and this arrangement becomes the third biological dimer geometry for MurI found | Mycobacterium tuberculosis | |
| 5.1.1.3 | additional information | inhibitor screening, compounds designed to target other glutamate racemases have been screened but do not inhibit mycobacterial MurI, suggesting that a different drug design effort will be needed to develop inhibitors. A distinct type of MurI dimer arrangement is observed in both structures, and this arrangement becomes the third biological dimer geometry for MurI found | Mycolicibacterium smegmatis |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.3 | 54200 | - |
recombinant enzyme, gel filtration | Mycobacterium tuberculosis |
| 5.1.1.3 | 59500 | - |
recombinant enzyme, gel filtration | Mycolicibacterium smegmatis |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.3 | L-glutamate | Mycobacterium tuberculosis | - |
D-glutamate | - |
r |  |
| 5.1.1.3 | L-glutamate | Mycolicibacterium smegmatis | - |
D-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | Mycobacterium tuberculosis ATCC 25618 / H37Rv | - |
D-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | Mycolicibacterium smegmatis ATCC 700084 / mc(2)155 | - |
D-glutamate | - |
r | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.1.1.3 | Mycobacterium tuberculosis | P9WPW9 | - |
- |
| 5.1.1.3 | Mycobacterium tuberculosis ATCC 25618 / H37Rv | P9WPW9 | - |
- |
| 5.1.1.3 | Mycolicibacterium smegmatis | A0R1X0 | - |
- |
| 5.1.1.3 | Mycolicibacterium smegmatis ATCC 700084 / mc(2)155 | A0R1X0 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.1.1.3 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag removal by TEV protease, and gel filtration, to over 95% purity | Mycobacterium tuberculosis |
| 5.1.1.3 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag removal by TEV protease, and gel filtration, to over 95% purity | Mycolicibacterium smegmatis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.3 | D-glutamate | D-Glu binding structure, overview | Mycobacterium tuberculosis | L-glutamate | - |
r | |
| 5.1.1.3 | D-glutamate | D-Glu binding structure, overview | Mycolicibacterium smegmatis | L-glutamate | - |
r | |
| 5.1.1.3 | D-glutamate | D-Glu binding structure, overview | Mycobacterium tuberculosis ATCC 25618 / H37Rv | L-glutamate | - |
r | |
| 5.1.1.3 | D-glutamate | D-Glu binding structure, overview | Mycolicibacterium smegmatis ATCC 700084 / mc(2)155 | L-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | - |
Mycobacterium tuberculosis | D-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | - |
Mycolicibacterium smegmatis | D-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | - |
Mycobacterium tuberculosis ATCC 25618 / H37Rv | D-glutamate | - |
r | |
| 5.1.1.3 | L-glutamate | - |
Mycolicibacterium smegmatis ATCC 700084 / mc(2)155 | D-glutamate | - |
r | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.1.1.3 | homodimer | 2 * 28900, about, sequence calcuation and analytical ultracentrifugation | Mycobacterium tuberculosis |
| 5.1.1.3 | homodimer | 2 * 29500, about, sequence calcuation and analytical ultracentrifugation | Mycolicibacterium smegmatis |
| 5.1.1.3 | More | the active site is located in a surface-exposed cleft that is formed at the interface of domains 1 and 2, subunit interaction analysis | Mycobacterium tuberculosis |
| 5.1.1.3 | More | the active site is located in a surface-exposed cleft that is formed at the interface of domains 1 and 2, subunit interaction analysis | Mycolicibacterium smegmatis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.1.1.3 | MurI | - |
Mycobacterium tuberculosis |
| 5.1.1.3 | MurI | - |
Mycolicibacterium smegmatis |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.1.1.3 | additional information | the mycobacterial MurI dimer is tightly associated, with a KD in the nanomolar range. The enzyme binds D- and L-glutamate specifically, but is inactive in solution unless the dimer interface is mutated. Enzyme structure analysis, active site structure and substrate binding structure, overview | Mycobacterium tuberculosis |
| 5.1.1.3 | additional information | the mycobacterial MurI dimer is tightly associated, with a KD in the nanomolar range. The enzyme binds D- and L-glutamate specifically, but is inactive in solution unless the dimer interface is mutated. Enzyme structure analysis, active site structure and substrate binding structure, overview | Mycolicibacterium smegmatis |
| 5.1.1.3 | physiological function | glutamate racemase (MurI) is responsible for providing D-glutamate for peptidoglycan biosynthesis in bacteria | Mycobacterium tuberculosis |
| 5.1.1.3 | physiological function | glutamate racemase (MurI) is responsible for providing D-glutamate for peptidoglycan biosynthesis in bacteria | Mycolicibacterium smegmatis |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
