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show all sequences of 1.1.1.214

Structure of conjugated polyketone reductase from Candida parapsilosis IFO 0708 reveals conformational changes for substrate recognition upon NADPH binding

Qin, H.M.; Yamamura, A.; Miyakawa, T.; Kataoka, M.; Nagai, T.; Kitamura, N.; Urano, N.; Maruoka, S.; Ohtsuka, J.; Nagata, K.; Shimizu, S.; Tanokura, M.; Appl. Microbiol. Biotechnol. 98, 243-249 (2013)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene cpr-c2, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3)
Candida parapsilosis
Crystallization (Commentary)
Crystallization
Organism
purified enzyme in apoform and in complex with NADPH, mixing of 0.001 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution consisting of 0.1 M Tris-HCl, pH 8.1, and 23% w/v PEG 3350 at 20°C, for the enzyme-NADPH complexed crystals, the protein in 20 mM Tris-HCl, pH 8.0, and 5 mM MADPH, is mixed with 0.1 M Tris–HCl, pH 7.4, and 256% w/v PEG 3350, X-ray diffraction structure determination and analysis at 1.70 A and 1.80 A resolution, respectively, molecular replacement method
Candida parapsilosis
Engineering
Amino acid exchange
Commentary
Organism
D58A
site-directed mutagenesis, the mutant shows 4.82% of wild-type activity
Candida parapsilosis
F299A
site-directed mutagenesis, the mutant shows 19.1% of wild-type activity
Candida parapsilosis
F300A
site-directed mutagenesis, the mutant shows 17.8% of wild-type activity
Candida parapsilosis
H125A
site-directed mutagenesis, the mutant shows 1.5% of wild-type activity
Candida parapsilosis
K264A
site-directed mutagenesis, the mutant shows 65.7% of wild-type activity
Candida parapsilosis
K28A
site-directed mutagenesis, the mutant shows 71.1% of wild-type activity
Candida parapsilosis
K30A
site-directed mutagenesis, the mutant shows 55.1% of wild-type activity
Candida parapsilosis
K88A
site-directed mutagenesis, inactive mutant
Candida parapsilosis
R267A
site-directed mutagenesis, the mutant shows 8.43% of wild-type activity
Candida parapsilosis
T27A
site-directed mutagenesis, the mutant shows 5.75% of wild-type activity
Candida parapsilosis
Y63A
site-directed mutagenesis, the mutant shows 0.17% of wild-type activity
Candida parapsilosis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(R)-pantolactone + NADP+
Candida parapsilosis
-
2-dehydropantolactone + NADPH + H+
-
-
r
(R)-pantolactone + NADP+
Candida parapsilosis IFO 0708
-
2-dehydropantolactone + NADPH + H+
-
-
r
2-dehydropantolactone + NADPH + H+
Candida parapsilosis
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
(R)-pantolactone + NADP+
-
-
r
2-dehydropantolactone + NADPH + H+
Candida parapsilosis IFO 0708
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
(R)-pantolactone + NADP+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Candida parapsilosis
Q76L36
-
-
Candida parapsilosis IFO 0708
Q76L36
-
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography, and tag cleavage by thrombin, followed by anion exchange chromatography, gel filtration, and dialysis
Candida parapsilosis
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
23.6
-
purified recombinant detagged wild-type enzyme, pH and temperature not specified in the publication
Candida parapsilosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-pantolactone + NADP+
-
724098
Candida parapsilosis
2-dehydropantolactone + NADPH + H+
-
-
-
r
(R)-pantolactone + NADP+
-
724098
Candida parapsilosis IFO 0708
2-dehydropantolactone + NADPH + H+
-
-
-
r
2-dehydropantolactone + NADPH + H+
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
724098
Candida parapsilosis
(R)-pantolactone + NADP+
-
-
-
r
2-dehydropantolactone + NADPH + H+
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
724098
Candida parapsilosis IFO 0708
(R)-pantolactone + NADP+
-
-
-
r
additional information
structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling
724098
Candida parapsilosis
?
-
-
-
-
additional information
structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling
724098
Candida parapsilosis IFO 0708
?
-
-
-
-
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
-
Candida parapsilosis
NADPH
-
Candida parapsilosis
Cloned(Commentary) (protein specific)
Commentary
Organism
gene cpr-c2, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3)
Candida parapsilosis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
-
Candida parapsilosis
NADPH
-
Candida parapsilosis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified enzyme in apoform and in complex with NADPH, mixing of 0.001 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution consisting of 0.1 M Tris-HCl, pH 8.1, and 23% w/v PEG 3350 at 20°C, for the enzyme-NADPH complexed crystals, the protein in 20 mM Tris-HCl, pH 8.0, and 5 mM MADPH, is mixed with 0.1 M Tris–HCl, pH 7.4, and 256% w/v PEG 3350, X-ray diffraction structure determination and analysis at 1.70 A and 1.80 A resolution, respectively, molecular replacement method
Candida parapsilosis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D58A
site-directed mutagenesis, the mutant shows 4.82% of wild-type activity
Candida parapsilosis
F299A
site-directed mutagenesis, the mutant shows 19.1% of wild-type activity
Candida parapsilosis
F300A
site-directed mutagenesis, the mutant shows 17.8% of wild-type activity
Candida parapsilosis
H125A
site-directed mutagenesis, the mutant shows 1.5% of wild-type activity
Candida parapsilosis
K264A
site-directed mutagenesis, the mutant shows 65.7% of wild-type activity
Candida parapsilosis
K28A
site-directed mutagenesis, the mutant shows 71.1% of wild-type activity
Candida parapsilosis
K30A
site-directed mutagenesis, the mutant shows 55.1% of wild-type activity
Candida parapsilosis
K88A
site-directed mutagenesis, inactive mutant
Candida parapsilosis
R267A
site-directed mutagenesis, the mutant shows 8.43% of wild-type activity
Candida parapsilosis
T27A
site-directed mutagenesis, the mutant shows 5.75% of wild-type activity
Candida parapsilosis
Y63A
site-directed mutagenesis, the mutant shows 0.17% of wild-type activity
Candida parapsilosis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(R)-pantolactone + NADP+
Candida parapsilosis
-
2-dehydropantolactone + NADPH + H+
-
-
r
(R)-pantolactone + NADP+
Candida parapsilosis IFO 0708
-
2-dehydropantolactone + NADPH + H+
-
-
r
2-dehydropantolactone + NADPH + H+
Candida parapsilosis
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
(R)-pantolactone + NADP+
-
-
r
2-dehydropantolactone + NADPH + H+
Candida parapsilosis IFO 0708
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
(R)-pantolactone + NADP+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography, and tag cleavage by thrombin, followed by anion exchange chromatography, gel filtration, and dialysis
Candida parapsilosis
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
23.6
-
purified recombinant detagged wild-type enzyme, pH and temperature not specified in the publication
Candida parapsilosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-pantolactone + NADP+
-
724098
Candida parapsilosis
2-dehydropantolactone + NADPH + H+
-
-
-
r
(R)-pantolactone + NADP+
-
724098
Candida parapsilosis IFO 0708
2-dehydropantolactone + NADPH + H+
-
-
-
r
2-dehydropantolactone + NADPH + H+
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
724098
Candida parapsilosis
(R)-pantolactone + NADP+
-
-
-
r
2-dehydropantolactone + NADPH + H+
the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner
724098
Candida parapsilosis IFO 0708
(R)-pantolactone + NADP+
-
-
-
r
additional information
structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling
724098
Candida parapsilosis
?
-
-
-
-
additional information
structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling
724098
Candida parapsilosis IFO 0708
?
-
-
-
-
General Information
General Information
Commentary
Organism
evolution
the conjugated polyketone reductase C2 (CPR-C2) from Candida parapsilosis IFO 0708 belongs to the aldo-keto reductase, AKR, superfamily
Candida parapsilosis
additional information
catalytic tetrad in the active site of CPR-C2/NADPH
Candida parapsilosis
General Information (protein specific)
General Information
Commentary
Organism
evolution
the conjugated polyketone reductase C2 (CPR-C2) from Candida parapsilosis IFO 0708 belongs to the aldo-keto reductase, AKR, superfamily
Candida parapsilosis
additional information
catalytic tetrad in the active site of CPR-C2/NADPH
Candida parapsilosis
Other publictions for EC 1.1.1.214
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)
724098
Qin
Structure of conjugated polyke ...
Candida parapsilosis, Candida parapsilosis IFO 0708
Appl. Microbiol. Biotechnol.
98
243-249
2013
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1
1
11
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4
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6
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1
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1
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6
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2
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1
2
1
11
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4
-
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1
-
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1
-
6
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
726465
Qin
Crystal structure of conjugate ...
Candida parapsilosis, Candida parapsilosis IFO 0708
Proteins
81
2059-2063
2013
-
-
-
1
-
-
-
-
-
-
1
4
-
4
-
-
-
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8
2
-
-
-
-
-
-
-
2
-
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-
-
-
2
1
-
-
-
-
-
-
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-
1
4
-
-
-
-
-
-
-
-
8
2
-
-
-
-
-
-
-
-
-
2
2
-
-
-
654311
Kataoka
Gene cloning and overexpressio ...
Candida parapsilosis, Candida parapsilosis IFO 0708
Appl. Microbiol. Biotechnol.
64
359-366
2004
-
1
1
-
-
-
1
-
-
-
2
2
-
11
-
-
-
-
-
-
2
-
4
1
1
-
-
-
1
1
-
1
-
-
-
-
1
1
1
-
-
-
-
1
-
-
-
-
2
2
-
-
-
-
-
-
2
-
4
1
1
-
-
-
1
1
-
-
-
-
-
-
-
-
347731
Julliard
-
Purification and characterizat ...
Spinacia oleracea
Bot. Acta
107
191-200
1994
-
-
-
-
-
-
2
-
1
-
1
1
-
1
-
-
1
-
-
-
1
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
1
-
1
1
-
-
-
1
-
-
1
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286105
Wilken
Ketopantoic acid and ketopanto ...
Escherichia coli
J. Biol. Chem.
250
2311-2314
1975
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-