Application | Comment | Organism |
---|---|---|
drug development | the enzyme IMPDH is a promising target for the treatment of Cryptosporidium infections | Cryptosporidium parvum |
Cloned (Comment) | Organism |
---|---|
recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) | Cryptosporidium parvum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme in complex with substrate IMP and inhibitor P131, sitting drop vapor diffusion method, mixing of 400 nl of 13.4 mg/ml protein in 20 mM HEPES, pH 8.0, 150 mM KCl, and 1.5 mM TCEP with 400 nl of reservoir solution containing 0.1 M bicine-NaOH, pH 9.0, and 20% PEG 6000, and equilibration against 0.134 ml of reservoir solution, at 16°C, X-ray diffraction structure determination and analysis at 2.053 A resolution, molecular replacement using the structure of CpIMPDH, PDB ID 3ffs, as search model, modelling | Cryptosporidium parvum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
N-(2-{3-[(1E)-N-(2-aminoethoxy)ethanimidoyl]phenyl}propan-2-yl)-N'-(4-chloro-3-nitrophenyl)urea | an inhibitor with in vivo anticryptosporidial activity, binding structure determination and analysi, P131 contains two aromatic groups, one of which interacts with the hypoxanthine ring of IMP, while the second interacts with the aromatic ring of a tyrosine in the adjacent subunit. In addition, the amine and NO2 moieties bind in hydrated cavities, forming water-mediated hydrogen bonds to the proteins. The oxime group of P131 lies across the hypoxanthine ring of IMP, while the N5 amine forms hydrogen bonds to the main chain carbonyl O atom of Gly212 and N3 of IMP in all of the subunits | Cryptosporidium parvum | |
N-(4-bromophenyl)-2-[2-(1,3-thiazol-2-yl)-1H-benzimidazol-1-yl]acetamide | - |
Cryptosporidium parvum | |
N-(4-chloro-3-methoxyphenyl)-2-(4-oxo[1]benzopyrano[4,3-c]pyrazol-1(4H)-yl)acetamide | - |
Cryptosporidium parvum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
38459 | - |
- |
Cryptosporidium parvum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | Cryptosporidium parvum | - |
XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | Cryptosporidium parvum Iowa II | - |
XMP + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cryptosporidium parvum | Q8T6T2 | - |
- |
Cryptosporidium parvum Iowa II | Q8T6T2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, His6-tag cleavage using recombinant TEV protease, and additional nickel affinity chromatography to remove the protease, the uncut protein and the affinity tag, followed by dialysis | Cryptosporidium parvum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | - |
Cryptosporidium parvum | XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | - |
Cryptosporidium parvum Iowa II | XMP + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 38459, sequence calculation | Cryptosporidium parvum |
More | IMPDH is a homotetramer with square-planar symmetry. The four active sites are located near the subunit interfaces. The IMP site is contained within a monomer, and the residues that contact IMP are strongly conserved among all IMPDHs | Cryptosporidium parvum |
Synonyms | Comment | Organism |
---|---|---|
IMPDH | - |
Cryptosporidium parvum |
inosine 5'-monophosphate dehydrogenase | - |
Cryptosporidium parvum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | cofactor-binding site, modelling, overview | Cryptosporidium parvum |
General Information | Comment | Organism |
---|---|---|
additional information | substrate-binding siteinvolving Cys219 and Met302-Gly303, modelling, overview | Cryptosporidium parvum |