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Literature summary for 1.1.1.317 extracted from

  • Sun, L.; Chen, Y.; Rajendran, C.; Mueller, U.; Panjikar, S.; Wang, M.; Mindnich, R.; Rosenthal, C.; Penning, T.M.; Stoeckigt, J.
    Crystal structure of perakine reductase, founding member of a novel aldo-keto reductase (AKR) subfamily that undergoes unique conformational changes during NADPH binding (2012), J. Biol. Chem., 287, 11213-11221.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression as N-terminal His6-tagged enzyme in Escherichia coli strain M15 Rauvolfia serpentina

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant methylated His6-tagged enzyme wild-type and mutant A213W, hanging drop vapor diffusion method, mixing of 0.002 ml of 5.5 mg/ml protein in 10 mM Tris-HCl buffer, pH 7.0, 1 mM DTT, 10 mM EDTA, with 0002 ml reservoir solution containing 25% v/v PEG 4000, 0.1 mM sodium citrate, pH 5.6, equilibration against 1 ml of reservoir solution at 20°C for 7 days, X-ray diffraction structure determination and analysis at 2.31 A and 1.77 A resolution, respectively Rauvolfia serpentina

Protein Variants

Protein Variants Comment Organism
A213W site-directed mutagenesis, cofactor-bound structure analysis Rauvolfia serpentina
additional information methylation of wild-type and mutant enzymes for improvement of the crystallization process Rauvolfia serpentina

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
perakine + NADPH + H+ Rauvolfia serpentina
-
raucaffrinoline + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Rauvolfia serpentina Q3L181
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
perakine + NADPH + H+
-
Rauvolfia serpentina raucaffrinoline + NADP+
-
?

Subunits

Subunits Comment Organism
More three-dimensional structure modeling of wild-type and mutant apo methylated enzymes with (alpha/beta)8-barrels with eight parallel beta-strands and eight alpha-helices typical for AKR superfamily members, overview Rauvolfia serpentina

Synonyms

Synonyms Comment Organism
AKR13D1
-
Rauvolfia serpentina

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Rauvolfia serpentina

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Rauvolfia serpentina

Cofactor

Cofactor Comment Organism Structure
NADPH dependent on, binding mode and structure, conformational changes caused by the cofactor binding, overview Rauvolfia serpentina

General Information

General Information Comment Organism
evolution the enzyme belongs to the AKR13D subfamily of the aldo-keto reductases, evolutionary relationship of AKR13D from Rauvolfia serpentina to annotated AKR families and subfamilies Rauvolfia serpentina
additional information the active site is formed by the catalytic tetrad Asp52, Tyr57, Lys84, and His126 at the center of the (alpha/beta)8-barrel structure. Upon NADPH binding, dramatic conformational changes and movements are observed: two additional beta-strands in the C terminus become ordered to form one alpha-helix, and a movement of up to 24 A occurs. This conformational change creates a large space that allows the binding of substrates of variable size for PR and enhances the enzyme activity Rauvolfia serpentina
physiological function perakine reductase catalyzes the NADPH-dependent reduction of the aldehyde perakine to yield the alcohol raucaffrinoline in the biosynthetic pathway of ajmaline in Rauvolfia, a key step in indole alkaloid biosynthesis Rauvolfia serpentina