Cloned (Comment) | Organism |
---|---|
gene TK1968, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta2 (DE3) pLysS | Thermococcus kodakarensis |
Crystallization (Comment) | Organism |
---|---|
purified enzyme in complex with CoA and 2-oxopantoate, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein and 1 mM CoA and 1 mM 2-oxopantoate with 0.001 ml of reservoir solution containing 100 mM Na acetate, pH 4.5, 20-25% v/v 2-methyl-2,4-pentanediol, and equilibration against 0.5 ml of reservoir solution, at 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.65 A resolution, modeling by molecular replacement method using N-terminal (1-165 residues) and C-terminal (171-309 residues) domains of Ec-KPR structure, PDB ID 2OFP, as separated search models, respectively | Thermococcus kodakarensis |
Protein Variants | Comment | Organism |
---|---|---|
C84A | site-directed mutagenesis, crystal structure analysis, overview | Thermococcus kodakarensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CoA | feedback inhibition, competitive with NADPH, the binding sites of CoA and NADP+ overlap, explaining the competitive manner of inhibition by CoA, binding structure analysis, overview | Thermococcus kodakarensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-pantoate + NADP+ | Thermococcus kodakarensis | - |
2-dehydropantoate + NADPH + H+ | - |
r | |
(R)-pantoate + NADP+ | Thermococcus kodakarensis ATCC BAA-918 | - |
2-dehydropantoate + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | Q5JGC2 | - |
- |
Thermococcus kodakarensis ATCC BAA-918 | Q5JGC2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta2 (DE3) pLysS by nickel affinity chromatography, avidin affinity chromatography, and dialysis | Thermococcus kodakarensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-pantoate + NADP+ | - |
Thermococcus kodakarensis | 2-dehydropantoate + NADPH + H+ | - |
r | |
(R)-pantoate + NADP+ | substrate binding structure analysis, overview | Thermococcus kodakarensis | 2-dehydropantoate + NADPH + H+ | - |
r | |
(R)-pantoate + NADP+ | - |
Thermococcus kodakarensis ATCC BAA-918 | 2-dehydropantoate + NADPH + H+ | - |
r | |
(R)-pantoate + NADP+ | substrate binding structure analysis, overview | Thermococcus kodakarensis ATCC BAA-918 | 2-dehydropantoate + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | enzyme structure analysis and comparisons, overview | Thermococcus kodakarensis |
Synonyms | Comment | Organism |
---|---|---|
ketopantoate reductase | - |
Thermococcus kodakarensis |
KPR | - |
Thermococcus kodakarensis |
Tk-KPR | - |
Thermococcus kodakarensis |
TK1968 | - |
Thermococcus kodakarensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
assay at | Thermococcus kodakarensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.4 | - |
assay at | Thermococcus kodakarensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | the binding sites of CoA and NADP+ overlap, explaining the competitive manner of inhibition by CoA, binding structure analysis, overview | Thermococcus kodakarensis |
General Information | Comment | Organism |
---|---|---|
additional information | CoA and 2-oxopantoate cooperatively trigger a conformational change from an open form to a closed enzyme form, structure analysis, overview | Thermococcus kodakarensis |
physiological function | ketopantoate reductase (KPR) catalyzes the NAD(P)H-dependent reduction of 2-oxopantoate to pantoate, and is a target of the feedback inhibition by CoA in archaea. Coenzyme A (CoA) plays essential roles in a variety of metabolic pathways in all three domains of life. The biosynthesis pathway of CoA is strictly regulated by feedback inhibition. In bacteria and eukaryotes, pantothenate kinase is the target of feedback inhibition by CoA | Thermococcus kodakarensis |