EC Number | Cloned (Comment) | Organism |
---|---|---|
5.4.99.9 | recombinant overexpression of active soluble His6-MBP-tagged MtUGM in Escherichia coli strain BL21(DE3) CodonPlus-RIL | Mycobacterium tuberculosis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
5.4.99.9 | purified detagged recombinant enzyme complexed with substrate UDP-Galp and inhibitors UDP, UDP-F4-Galf, and UDP-F4-Galp, hanging drop vapour diffusion method, mixing of 0.0012 ml of protein solution containing 6.5 mg/ml enzyme in 25 mM Tris, pH 7.5, 500 mM NaCl, and, added prior to crystallization, 20 mM sodium dithionite (final concentration), with 0.0012 ml of reservoir solution containing 20% PEG 3350, 0.1 M Bis-Tris pH 5.5, and additives, 1-4 weeks, X-ray diffraction structure determination and analysis at 2.25-2.60 A resolution | Mycobacterium tuberculosis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
5.4.99.9 | P306R | site-directed mutagenesis, the mutant shows similar kinetics compared to wild-type. Pro306 is located on the solvent-exposed loop (His300-Lys309) connecting the small helix nu3 and beta strand beta14 of the beta-sheet domain, over 25 A from the FAD. The Cdelta atom of Pro306 is 3.5 A from the main-chain oxygen of Thr53, located on the small sharp turn (Glu52-Gly56) connecting beta strands beta3 and beta4.The Pro306Arg mutation releases this clash and results in a 1 A shift in the position of the two solvent-exposed loops without affecting the position of side chains and interaction with the protein. Arg306 forms a salt bridge with the side chain of Asp308 and the main-chain oxygen of Gln54 and replaces a salt bridge formed by Lys309. The Lys309 side chain has rotated and forms a new salt bridge with Gln54. In addition, Lys309 is involved in crystal contacts with the side chain of Asp202. The Pro306Ala mutation therefore may be stabilizing the two loops and promoting the crystallization of MtUGM complex structures | Mycobacterium tuberculosis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
5.4.99.9 | additional information | dissociation constants for the inhibitors by saturation transfer difference (STD) NMR spectroscopy measurements | Mycobacterium tuberculosis | |
5.4.99.9 | UDP | enzyme binding structure analysis and binding mode, overview | Mycobacterium tuberculosis | |
5.4.99.9 | UDP-2,3-dideoxy-2,2,3,3-tetrafluoro-alpha-D-galactofuranose | UDP-F4-Galf, enzyme binding structure analysis and binding mode, overview | Mycobacterium tuberculosis | |
5.4.99.9 | UDP-2,3-dideoxy-2,2,3,3-tetrafluoro-alpha-D-galactopyranose | UDP-F4-Galp, enzyme binding structure analysis and binding mode, overview | Mycobacterium tuberculosis | |
5.4.99.9 | UDP-3-deoxy-3-fluoro-alpha-D-galactofuranose | - |
Mycobacterium tuberculosis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.4.99.9 | 0.023 | - |
UDP-2,3-dideoxy-2,2,3,3-tetrafluoro-alpha-D-galactopyranose | pH 7.5, 37°C, recombinant enzyme | Mycobacterium tuberculosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.4.99.9 | UDP-alpha-D-galactopyranose | Mycobacterium tuberculosis | - |
UDP-alpha-D-galactofuranose | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.4.99.9 | Mycobacterium tuberculosis | P9WIQ1 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.4.99.9 | recombinant soluble His6-MBP-tagged MtUGM from Escherichia coli strain BL21(DE3) CodonPlus-RIL by amylose affinity chromatography, tag cleavage by TEV protease, and elimination of residual tag and tagged enzyme by nickel affinity chromatography | Mycobacterium tuberculosis |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
5.4.99.9 | UDP-alpha-D-galactopyranose = UDP-alpha-D-galactofuranose | enzyme-substrate binding structure and mechanism | Mycobacterium tuberculosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.4.99.9 | additional information | analysis of the structural basis of ligand binding to UDP-galactopyranose mutase from Mycobacterium tuberculosis using substrate and tetrafluorinated substrate analogues, overview | Mycobacterium tuberculosis | ? | - |
? | |
5.4.99.9 | UDP-2,3-dideoxy-2,2,3,3-tetrafluoro-alpha-D-galactopyranose | - |
Mycobacterium tuberculosis | UDP-2,3-dideoxy-2,2,3,3-tetrafluoro-alpha-D-galactofuranose | - |
r | |
5.4.99.9 | UDP-alpha-D-galactopyranose | - |
Mycobacterium tuberculosis | UDP-alpha-D-galactofuranose | - |
r | |
5.4.99.9 | UDP-alpha-D-galactopyranose | substrate binding structure of wild-type and mutant P306R, three-dimensional structure of the reduced MtUGM:UDP-Galp dimer, detailed overview. Substrate binding induces local changes in MtUGM active site | Mycobacterium tuberculosis | UDP-alpha-D-galactofuranose | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
5.4.99.9 | dimer | - |
Mycobacterium tuberculosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.4.99.9 | MtUGM | - |
Mycobacterium tuberculosis |
5.4.99.9 | UGM | - |
Mycobacterium tuberculosis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
5.4.99.9 | 37 | - |
assay at | Mycobacterium tuberculosis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.4.99.9 | 7.5 | - |
assay at | Mycobacterium tuberculosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
5.4.99.9 | FAD | enzyme interaction mechanism, overview | Mycobacterium tuberculosis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
5.4.99.9 | physiological function | UDP-galactopyranose mutase (UGM) is a flavin-containing enzyme that catalyzes the reversible conversion of UDP-galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf) and plays a key role in the biosynthesis of the mycobacterial cell wall galactofuran. Substrate binding induces local changes in MtUGM active site | Mycobacterium tuberculosis |