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show all sequences of 1.1.1.317

Crystal structure of perakine reductase, founding member of a novel aldo-keto reductase (AKR) subfamily that undergoes unique conformational changes during NADPH binding

Sun, L.; Chen, Y.; Rajendran, C.; Mueller, U.; Panjikar, S.; Wang, M.; Mindnich, R.; Rosenthal, C.; Penning, T.M.; Stoeckigt, J.; J. Biol. Chem. 287, 11213-11221 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression as N-terminal His6-tagged enzyme in Escherichia coli strain M15
Rauvolfia serpentina
Crystallization (Commentary)
Crystallization
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
Engineering
Amino acid exchange
Commentary
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
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
perakine + NADPH + H+
Rauvolfia serpentina
-
raucaffrinoline + NADP+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Rauvolfia serpentina
Q3L181
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
perakine + NADPH + H+
-
725466
Rauvolfia serpentina
raucaffrinoline + NADP+
-
-
-
?
Subunits
Subunits
Commentary
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
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Rauvolfia serpentina
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Rauvolfia serpentina
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
dependent on, binding mode and structure, conformational changes caused by the cofactor binding, overview
Rauvolfia serpentina
Cloned(Commentary) (protein specific)
Commentary
Organism
expression as N-terminal His6-tagged enzyme in Escherichia coli strain M15
Rauvolfia serpentina
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
dependent on, binding mode and structure, conformational changes caused by the cofactor binding, overview
Rauvolfia serpentina
Crystallization (Commentary) (protein specific)
Crystallization
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
Engineering (protein specific)
Amino acid exchange
Commentary
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) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
perakine + NADPH + H+
Rauvolfia serpentina
-
raucaffrinoline + NADP+
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
perakine + NADPH + H+
-
725466
Rauvolfia serpentina
raucaffrinoline + NADP+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
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
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Rauvolfia serpentina
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Rauvolfia serpentina
General Information
General Information
Commentary
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
General Information (protein specific)
General Information
Commentary
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
Other publictions for EC 1.1.1.317
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
725466
Sun
Crystal structure of perakine ...
Rauvolfia serpentina
J. Biol. Chem.
287
11213-11221
2012
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1
1
2
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1
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2
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-
-
-
-
-
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1
1
1
-
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1
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1
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1
1
1
2
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1
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1
1
1
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-
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1
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3
3
-
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-
716558
Sun
Purification, cloning, functio ...
Rauvolfia serpentina
Plant Mol. Biol.
67
455-467
2008
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1
-
5
-
-
2
-
-
2
1
-
5
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1
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-
-
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4
1
2
-
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2
2
1
-
1
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1
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1
1
-
5
-
-
-
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2
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2
1
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1
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4
1
2
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2
2
1
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1
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-
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2
2
713623
Rosenthal
Expression, purification, crys ...
Rauvolfia serpentina
Acta Crystallogr. Sect. F
62
1286-1289
2006
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1
1
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1
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1
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1
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1
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1
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1
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1
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