Cloned (Comment) | Organism |
---|---|
gene panE, expression of His6-tagged wild-type and mutant enzymes | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified enzyme with bound NADP+, hanging drop vapour diffusion method, 10-15 mg/ml protein at 4°C is mixed with ketopantoate and NADP+ in a ratio of 5:1 and 2:1, respectively, in 0.1 M sodium acetate, pH 4.0-5.0, with 10%2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.1 A resolution, ternary complex modelling | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D248A | site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain | Escherichia coli |
E210A | site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain | Escherichia coli |
E256A | site-directed mutagenesis, nearly inactive mutant, 2600fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain | Escherichia coli |
K176A | site-directed mutagenesis, nearly inactive mutant, 78000fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain | Escherichia coli |
K72A | site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain | Escherichia coli |
N98A | site-directed mutagenesis, nearly inactive mutant, 4000fold reduced catalytic efficiency, no complementation of a panE knockout mutant strain | Escherichia coli |
S244A | site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics and thermodynamics, wild-type enzyme, overview | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ketopantoate + NADPH | Escherichia coli | the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA | pantoate + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9J4 | gene panE | - |
Purification (Comment) | Organism |
---|---|
native wild-type enzyme by anion exchange and adsorption chromatography, and gel filtration, recombinant His6-tagged enzyme by nickel affinity chromatography | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(R)-pantoate + NADP+ = 2-dehydropantoate + NADPH + H+ | molecular catalytic mechanism, substrate and cofactor binding, Asn98, Glu256, and Lys176 are essential, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ketopantoate + NADPH | the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA | Escherichia coli | pantoate + NADP+ | - |
r | |
ketopantoate + NADPH | substrate binding structure and thermodynamics | Escherichia coli | pantoate + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ketopantoate reductase | - |
Escherichia coli |
KPR | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | cofactor binding structure and thermodynamics, the cofactor is bound in the active site cleft between the N-terminal Rossmann-fold domain and the C-terminal alpha-helical domain | Escherichia coli | |
NADPH | cofactor binding structure and thermodynamics | Escherichia coli |