Application | Comment | Organism |
---|---|---|
drug development | inosine 5'-monophosphate dehydrogenase 2 (IMPDH2) from Mycobacterium tuberculosis is an attractive drug target | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
gene guaB2, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant MtbIMPDH2DELTACBS enzyme mutant in apofomr or complexed with IMP/NAD+, or with inhibitors MAD1/IMP, P41/IMP, or Q67/IMP, the crystallization conditions are for the apoform mutant 0.1 M sodium/potassium phosphate, pH 6.2, 25% 1,2-propandiol, 10% glycerol, 16°C, for the MAD1/IMP-enzyme mutant complex 0.1 M sodium/potassium phosphate, pH 6.2, 25% 1,2-propandiol, 10% glycerol, 16°C, for the P41/IMP-enzyme mutant complex 0.3 M magnesium formate, 0.1 M Tris, pH 8.5, 16°C, for the Q67/IMP-enzyme mutant complex 0.1 M sodium/potassium phosphate, pH 6.2, 25% 1,2-propandiol, 10% glycerol, 16°C, and for the NAD+/IMP-enzyme mutant complex 0.4 M magnesium formate, 0.1M Tris-HCl, pH 8.5, 20% sucrose, 16°C, crystals are soaked with 200 mM ligand solutions, X-ray diffraction structure determination and analysis at 1.60-2.00 A resolution. Analysis of the crystal structure of the enzyme mutant in complex with XMP and NAD+, detailed overview | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | deletion of IMPDH CBS domain of MtbIMPDH2, residues E126-R252 are replaced with a GG linker. The MtbIMPDH2DELTACBS mutant shows significantly improved solubility and crystallizability properties compared to the wild-type enzyme, with the steady state kinetic parameters comparable to those reported for the wild-type enzyme | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(2R)-2-phenyl-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
(2S)-2-(2,3-dichlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
(2S)-2-(2,3-dichlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-6-yl]propanamide | - |
Mycobacterium tuberculosis | |
(2S)-2-(2,3-dimethoxyphenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
(2S)-2-(2-chloro-3-nitrophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
(2S)-2-(2-chlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
2-(2,4-dichlorophenoxy)-N-[2-(pyridin-2-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
2-(2-chlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
2-(4-methoxyphenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
2-chloro-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]-N,N-dimethylbenzamide | - |
Mycobacterium tuberculosis | |
2-chloro-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]benzamide | - |
Mycobacterium tuberculosis | |
2-chloro-N,N-diethyl-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]benzamide | - |
Mycobacterium tuberculosis | |
2-chloro-N,N-dimethyl-5-[([2-[3-(prop-1-en-2-yl)phenyl]propan-2-yl]carbamoyl)amino]benzamide | noncompetitive inhibitor, that has similar affinity to both E-IMP and E-XMP, enzyme binding structure analysis, overview | Mycobacterium tuberculosis | |
2-phenoxy-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | - |
Mycobacterium tuberculosis | |
5'-O-([1-[(2E)-4-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-dihydro-2-benzofuran-5-yl)-2-methylbut-2-en-1-yl]-1H-1,2,3-triazol-4-yl]methyl)adenosine | an uncompetitive inhibitor with a strong preference for the E-XMP reaction intermediate, enzyme binding structure analysis, overview | Mycobacterium tuberculosis | |
additional information | IMPDH has multiple active site states that can be targeted for inhibitor design, and both the IMP/XMP and cofactor binding sites are exploited for that purpose, inhibitor screening of compounds with antitubercular activity, overview. Mechanism of MtbIMPDH2DELTACBS enzyme mutant inhibition | Mycobacterium tuberculosis | |
N-[4-chloro-3-(morpholine-4-carbonyl)phenyl]-N'-(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)urea | - |
Mycobacterium tuberculosis | |
N2-(2,3-dichlorophenyl)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]-L-alaninamide | a competitive inhibitor versus NAD+, that has a strong preference for E-IMP, enzyme binding structure analysis, overview | Mycobacterium tuberculosis | |
NAD+ | low substrate inhibition | Mycobacterium tuberculosis | |
N~2~-(2,3-dichlorophenyl)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]alaninamide | - |
Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.041 | - |
IMP | recombinant MtbIMPDH2DELTACBS mutant enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.078 | - |
IMP | recombinant wild-type enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.58 | - |
NAD+ | recombinant MtbIMPDH2DELTACBS mutant enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
1.005 | - |
NAD+ | recombinant wild-type enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | Mycobacterium tuberculosis | - |
XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | Mycobacterium tuberculosis H37Rv | - |
XMP + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WKI7 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WKI7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Mycobacterium tuberculosis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
IMP + NAD+ + H2O = XMP + NADH + H+ | the enzymatic mechanism of IMPDH consists two steps, a dehydrogenase and a hydrolase reaction. Upon binding of IMP and NAD+ cofactor, a thioimidate enzyme-substrate adduct, E-XMP*, is formed via a covalent bond to the catalytic C341 with concurrent production of NADH. Hydride transfer occurs to the pro-S position with the cofactor in the anti-conformation. The cofactor is released and an active site mobile flap moves into the NAD+ site and facilitates E-XMP* hydrolysis with a conserved R443 acting as a general base. The enzyme has two distinct conformations, an open form for the dehydrogenase reaction and a closed form for E-XMP* hydrolysis | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | - |
Mycobacterium tuberculosis | XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | - |
Mycobacterium tuberculosis H37Rv | XMP + NADH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
guaB2 | - |
Mycobacterium tuberculosis |
IMPDH2 | - |
Mycobacterium tuberculosis |
inosine 5-monophosphate dehydrogenase 2 | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.53 | - |
NAD+ | recombinant wild-type enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.53 | - |
IMP | recombinant wild-type enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.57 | - |
NAD+ | recombinant MtbIMPDH2DELTACBS mutant enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.57 | - |
IMP | recombinant MtbIMPDH2DELTACBS mutant enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | dependent on, binding structure to CBS domain-deleted enzyme mutant of MtbIMPDH2DELTACBS | Mycobacterium tuberculosis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000013 | - |
2-chloro-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]-N,N-dimethylbenzamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000014 | - |
N2-(2,3-dichlorophenyl)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]-L-alaninamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000017 | - |
2-chloro-N,N-dimethyl-5-[([2-[3-(prop-1-en-2-yl)phenyl]propan-2-yl]carbamoyl)amino]benzamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000035 | - |
2-chloro-N,N-diethyl-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]benzamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000037 | - |
N-[4-chloro-3-(morpholine-4-carbonyl)phenyl]-N'-(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)urea | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00004 | - |
(2S)-2-(2,3-dichlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-6-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00004 | - |
N~2~-(2,3-dichlorophenyl)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]alaninamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000076 | - |
(2S)-2-(2,3-dichlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.0001 | - |
(2S)-2-(2-chlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00013 | - |
(2S)-2-(2-chloro-3-nitrophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00015 | - |
2-(4-methoxyphenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.000158 | - |
2-chloro-5-[[(2-[3-[(1Z)-N-hydroxyethanimidoyl]phenyl]propan-2-yl)carbamoyl]amino]benzamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00024 | - |
2-(2-chlorophenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00044 | - |
(2R)-2-phenyl-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00062 | - |
(2S)-2-(2,3-dimethoxyphenoxy)-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00065 | - |
2-phenoxy-N-[2-(pyridin-4-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.00158 | - |
5'-O-([1-[(2E)-4-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-dihydro-2-benzofuran-5-yl)-2-methylbut-2-en-1-yl]-1H-1,2,3-triazol-4-yl]methyl)adenosine | pH 8.0, 22°C | Mycobacterium tuberculosis | |
0.002 | - |
2-(2,4-dichlorophenoxy)-N-[2-(pyridin-2-yl)-1,3-benzoxazol-5-yl]propanamide | pH 8.0, 22°C | Mycobacterium tuberculosis | |
5 | - |
NAD+ | recombinant wild-type enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis | |
16 | - |
NAD+ | recombinant MtbIMPDH2DELTACBS mutant enzyme, pH 8.0, 22°C | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
additional information | determination of XMP and NAD+ substrate binding structures, overview. Active site structures of wild-type apo-enzyme and of CBS domain-deleted enzyme mutant MtbIMPDH2DELTACBS in apoform | Mycobacterium tuberculosis |
physiological function | the enzyme IMPDH2 catalyzes the conversion of inosine 5'-monophosphate into xanthosine 5'-monophosphate with the concomitant reduction of NAD+ to NADH and controls flux into the guanine nucleotide pool | Mycobacterium tuberculosis |