Cloned (Comment) | Organism |
---|---|
expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21 (DE3) pLysS | Coleus scutellarioides |
Crystallization (Comment) | Organism |
---|---|
apoenzyme and enzyme in complex with NADP+, sitting drop vapour diffusion method, mixing of 0.001 ml of 6 mg/ml protein or 15 mg/ml protein with 2 mM NADP+ in 50 mM Tris-HCl, pH 7.0, with 0.001 ml of well solution containing 30% 2-methyl-2,4-pentanediol, 0.2 M NaCl, 0.1 M Tris-HCl, pH 7.5 at 4°C, or 20% PEG 1000, 0.1 M imidazole, pH 7.5, 4 mM 4-hydroxyphenylpyruvate, 0.2 mM DTT at 26°C, leading to one-crystal clusters or tetragonal bipyramid-shaped crystals, X-ray diffraction structure determination and analysis at 1.47 A and 2.2 A, respectively, modeling | Coleus scutellarioides |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34113 | - |
2 * 34113, sequence calculation and crystal structure | Coleus scutellarioides |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-(4-hydroxyphenyl)pyruvate + NADPH + H+ | Coleus scutellarioides | - |
3-(4-hydroxyphenyl)lactate + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Coleus scutellarioides | Q65CJ7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity and 2',5'-ADP affinity chromatography and dialysis | Coleus scutellarioides |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell suspension culture | - |
Coleus scutellarioides | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3,4-dihydroxyphenylpyruvate + NADH + H+ | - |
Coleus scutellarioides | 3-(3,4-dihydroxyphenyl)lactate + NAD+ | - |
r | |
3,4-dihydroxyphenylpyruvate + NADPH + H+ | - |
Coleus scutellarioides | 3-(3,4-dihydroxyphenyl)lactate + NADP+ | - |
r | |
3-(4-hydroxyphenyl)pyruvate + NADH + H+ | - |
Coleus scutellarioides | 3-(4-hydroxyphenyl)lactate + NAD+ | - |
r | |
3-(4-hydroxyphenyl)pyruvate + NADPH + H+ | - |
Coleus scutellarioides | 3-(4-hydroxyphenyl)lactate + NADP+ | - |
? | |
3-(4-hydroxyphenyl)pyruvate + NADPH + H+ | substrate bindiing structure, docking study, overview | Coleus scutellarioides | 3-(4-hydroxyphenyl)lactate + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 34113, sequence calculation and crystal structure | Coleus scutellarioides |
More | molecular docking study, overview | Coleus scutellarioides |
Synonyms | Comment | Organism |
---|---|---|
H(P)PR | - |
Coleus scutellarioides |
HPPR | - |
Coleus scutellarioides |
hydroxy(phenyl)pyruvate reductase | - |
Coleus scutellarioides |
hydroxyphenylpyruvate reductase | - |
Coleus scutellarioides |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | NADH as well as NADPH can serve as the electron donor | Coleus scutellarioides | |
NADP+ | - |
Coleus scutellarioides | |
NADPH | preferred cofactor compared to NAD+, forms three hydrogen bonds to the protein via N41 to Asp256 OD2 and Ile230 O and O40 to His279 NE2 | Coleus scutellarioides |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the family of D-isomer-specific 2-hydroxyacid dehydrogenases | Coleus scutellarioides |
additional information | movement of the two domains after cosubstrate binding in order to close the inter-domain cleft for catalysis. The amino acids participating in the contacts are Leu205, Arg232 and His279 from the cosubstrate-binding domain and Ser53, Gly77 and Asp79 from the substrate-binding domain. The active site of H(P)PR is formed by the amino-acid residues Arg232 and His279, active site structure, overview | Coleus scutellarioides |