Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally His6-tagged or GSG-tagged enzyme | Corynebacterium diphtheriae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His6-tagged enzyme in complex with citrate and GSG-tagged enzyme in complex with UDP, by hanging drop vapor diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris, pH 7.0, with 0.002 ml of reservoir solution containing for the citrate-complexed enzyme 100 mM sodium citrate, pH 5.6, 15% isopropanol, and 16-18% PEG 5000 MME, for 3-4 days, and containing for the UDP-complexed enzyme 100 mM Bis-Tris, pH 5.5, 200 mM lithium sulfate, and 21% PEG 3350, for 1-2 weeks, X-ray diffraction structure determination and analysis at 1.95-2.35 A resolution, molecular replacement using KpUGM monomer structure in an open conformation (PDB ID 2BI7) as template for the citrate-complexed enzyme crystals and the structure of Mycobacterium smegmatis UGM (MsUGM) in a closed conformation (PDB ID 5EQD) for the UDP-complexed enzyme crystals, structure comparisons, overview | Corynebacterium diphtheriae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-((4-(3,4-dichlorophenyl)thiazol-2-yl)amino)-3-(4-iodophenyl)propanoic acid | - |
Corynebacterium diphtheriae | |
3-(4-bromobenzyl)-6-(4-chlorophenyl)-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine-7-carboxylic acid | - |
Corynebacterium diphtheriae | |
6-(4-chlorophenyl)-3-(thiophen-2-yl)-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine-7-carboxylic acid | - |
Corynebacterium diphtheriae | |
additional information | the enzyme can be inhibited by 2-aminothiazoles, e.g. 2-((4-(3,4-dichlorophenyl)thiazol-2-yl)amino)-3-(4-iodophenyl)propanoic acid, or triazolothiadiazines, such as 6-(4-chlorophenyl)-3-(thiophen-2-yl)-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine-7-carboxylic acid or 3-(4-bromobenzyl)-6-(4-chlorophenyl)-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine-7-carboxylic acid | Corynebacterium diphtheriae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
citrate | the citrate ion is identified on the basis of its shape and bonding partners and is located in the active site cavity adjacent to the FAD cofactor. Citrate is anionic like the UDP-alpha-D-galactopyranose substrate. The citrate ion forms salt bridges with UGM residues R288 and H290 and numerous ordered water molecules, thereby participating in a hydrogen-bonding network with the active site residues | Corynebacterium diphtheriae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-galactopyranose | Corynebacterium diphtheriae | - |
UDP-alpha-D-galactofuranose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium diphtheriae | Q6NER4 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant expression of His6-tagged or GSG-tagged enzyme by nickel affinity chromatography and gel filtration | Corynebacterium diphtheriae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-galactopyranose | - |
Corynebacterium diphtheriae | UDP-alpha-D-galactofuranose | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | structure of dimeric recombinant GSG-CdUGM | Corynebacterium diphtheriae |
Synonyms | Comment | Organism |
---|---|---|
CdUGM | - |
Corynebacterium diphtheriae |
Glf | - |
Corynebacterium diphtheriae |
UGM | - |
Corynebacterium diphtheriae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | tightly bound to the enzyme | Corynebacterium diphtheriae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0049 | - |
2-((4-(3,4-dichlorophenyl)thiazol-2-yl)amino)-3-(4-iodophenyl)propanoic acid | pH 7.0, temperature not specified in the publication | Corynebacterium diphtheriae |
General Information | Comment | Organism |
---|---|---|
additional information | active site structure, the citrate ion forms salt bridges with UGM residues R288 and H290 and numerous ordered water molecules, thereby participating in a hydrogen-bonding network with the active site residues. A strong density feature is observed bridging the citrate ion and the oxidized FAD | Corynebacterium diphtheriae |