Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.205 extracted from

  • Kim, Y.; Makowska-Grzyska, M.; Gorla, S.K.; Gollapalli, D.R.; Cuny, G.D.; Joachimiak, A.; Hedstrom, L.
    Structure of Cryptosporidium IMP dehydrogenase bound to an inhibitor with in vivo antiparasitic activity (2015), Acta Crystallogr. Sect. F, 71, 531-538.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
drug development the enzyme IMPDH is a promising target for the treatment of Cryptosporidium infections Cryptosporidium parvum

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Cryptosporidium parvum

Crystallization (Commentary)

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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
38459
-
-
Cryptosporidium parvum

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Cryptosporidium parvum Q8T6T2
-
-
Cryptosporidium parvum Iowa II Q8T6T2
-
-

Purification (Commentary)

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 and Products (Substrate)

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

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

Synonyms Comment Organism
IMPDH
-
Cryptosporidium parvum
inosine 5'-monophosphate dehydrogenase
-
Cryptosporidium parvum

Cofactor

Cofactor Comment Organism Structure
NAD+ cofactor-binding site, modelling, overview Cryptosporidium parvum

General Information

General Information Comment Organism
additional information substrate-binding siteinvolving Cys219 and Met302-Gly303, modelling, overview Cryptosporidium parvum