BRENDA - Enzyme Database show
show all sequences of 2.7.1.86

Structural determinants of discrimination of NAD+ from NADH in yeast mitochondrial NADH kinase Pos5

Ando, T.; Ohashi, K.; Ochiai, A.; Mikami, B.; Kawai, S.; Murata, K.; J. Biol. Chem. 286, 29984-29992 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli RosettaBlue(DE3) cells
Saccharomyces cerevisiae
Crystallization (Commentary)
Crystallization
Organism
Pos5 complexed with NADH, sitting drop vapor diffusion method, using 15% (v/v) 2-methyl 2,4-pentanediol, 5% (w/v) polyethylene glycol 4000 and 100 mM imidazole-HCl, pH 8.0, at 20°C
Saccharomyces cerevisiae
Engineering
Amino acid exchange
Commentary
Organism
H231D
the mutant exhibits no activity towards NAD+ and low activity for NADH
Saccharomyces cerevisiae
R293H
the mutation reduces the ratio of NADH kinase activity to NAD kinase activity from 8.6 to 2.1
Saccharomyces cerevisiae
R293H/H231D
inactive
Saccharomyces cerevisiae
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.032
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
0.19
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
1.3
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
4.5
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
mitochondrion
-
Saccharomyces cerevisiae
5739
-
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46285
-
2 * 46285, calculated from amino acid sequence
Saccharomyces cerevisiae
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Saccharomyces cerevisiae
Q06892
-
-
Purification (Commentary)
Commentary
Organism
Ni-NTA agarose column chromatography and Superdex 200 gel filtration
Saccharomyces cerevisiae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + NAD+
the enzyme has much higher NADH kinase than NAD kinase activity
722719
Saccharomyces cerevisiae
ADP + NADP+
-
-
-
?
ATP + NADH
the enzyme has much higher NADH kinase than NAD kinase activity
722719
Saccharomyces cerevisiae
ADP + NADPH
-
-
-
?
Subunits
Subunits
Commentary
Organism
homodimer
2 * 46285, calculated from amino acid sequence
Saccharomyces cerevisiae
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.2
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
7.4
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
7.7
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
16.1
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
optimum pH for NAD kinase activity of Pos5
Saccharomyces cerevisiae
9.5
-
optimum pH for NADH kinase activity of Pos5
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli RosettaBlue(DE3) cells
Saccharomyces cerevisiae
Crystallization (Commentary) (protein specific)
Crystallization
Organism
Pos5 complexed with NADH, sitting drop vapor diffusion method, using 15% (v/v) 2-methyl 2,4-pentanediol, 5% (w/v) polyethylene glycol 4000 and 100 mM imidazole-HCl, pH 8.0, at 20°C
Saccharomyces cerevisiae
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
H231D
the mutant exhibits no activity towards NAD+ and low activity for NADH
Saccharomyces cerevisiae
R293H
the mutation reduces the ratio of NADH kinase activity to NAD kinase activity from 8.6 to 2.1
Saccharomyces cerevisiae
R293H/H231D
inactive
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.032
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
0.19
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
1.3
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
4.5
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
mitochondrion
-
Saccharomyces cerevisiae
5739
-
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46285
-
2 * 46285, calculated from amino acid sequence
Saccharomyces cerevisiae
Purification (Commentary) (protein specific)
Commentary
Organism
Ni-NTA agarose column chromatography and Superdex 200 gel filtration
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + NAD+
the enzyme has much higher NADH kinase than NAD kinase activity
722719
Saccharomyces cerevisiae
ADP + NADP+
-
-
-
?
ATP + NADH
the enzyme has much higher NADH kinase than NAD kinase activity
722719
Saccharomyces cerevisiae
ADP + NADPH
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 46285, calculated from amino acid sequence
Saccharomyces cerevisiae
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.2
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
7.4
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
7.7
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
16.1
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
optimum pH for NAD kinase activity of Pos5
Saccharomyces cerevisiae
9.5
-
optimum pH for NADH kinase activity of Pos5
Saccharomyces cerevisiae
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.6
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
4.8
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
85
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
241
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.6
-
NAD+
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
4.8
-
NAD+
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
85
-
NADH
wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
241
-
NADH
mutant enzyme R293H, in 100 mM Tris-HCl, pH 8.0, at 37°C
Saccharomyces cerevisiae
Other publictions for EC 2.7.1.86
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)
721390
Lee
Effects of NADH kinase on NADP ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae CEN.PK2-1D
Appl. Microbiol. Biotechnol.
97
1561-1569
2013
-
-
1
-
-
-
-
-
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1
-
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4
-
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2
1
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1
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1
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2
1
-
-
-
-
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-
-
-
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-
-
-
721392
Lee
Engineering of NADPH regenerat ...
Saccharomyces cerevisiae
Appl. Microbiol. Biotechnol.
97
2761-2772
2013
-
-
1
-
-
-
-
-
1
-
-
-
-
4
-
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-
-
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-
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1
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1
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1
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1
-
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-
-
-
-
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-
-
-
-
-
-
721139
Shi
Role of mitochondrial NADH kin ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4742
Acta Biochim. Biophys. Sin.
43
989-995
2011
-
-
-
-
-
-
-
-
2
-
-
-
-
24
-
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2
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2
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-
2
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
722719
Ando
Structural determinants of dis ...
Saccharomyces cerevisiae
J. Biol. Chem.
286
29984-29992
2011
-
-
1
1
3
-
-
4
2
-
1
-
-
3
-
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1
-
-
-
-
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2
1
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4
2
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1
-
1
3
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4
2
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1
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1
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2
1
-
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4
2
-
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-
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-
4
4
706395
Waller
Subcellular and tissue localiz ...
Arabidopsis thaliana
Planta
231
305-317
2010
-
-
1
-
-
-
-
-
1
-
-
-
-
3
-
-
-
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3
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1
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1
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1
1
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1
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3
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1
-
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-
-
-
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-
-
1
-
-
1
-
-
722693
Pain
Mitochondrial NADH kinase, Pos ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4741
J. Biol. Chem.
285
39409-39424
2010
-
-
1
-
-
-
-
-
2
-
-
-
-
59
-
-
1
-
-
-
-
-
2
-
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-
-
-
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-
-
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1
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2
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1
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-
-
2
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
695829
Hou
Impact of overexpressing NADH ...
Saccharomyces cerevisiae
Appl. Microbiol. Biotechnol.
82
909-919
2009
-
-
-
-
-
-
-
-
2
-
-
1
-
4
-
-
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-
-
-
-
-
4
-
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-
-
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2
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3
-
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3
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1
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4
-
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-
-
-
-
-
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705593
Stuart
Transcriptional response to mi ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae YPH925
Mitochondrion
9
211-221
2009
-
-
-
-
-
-
-
-
2
-
-
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3
-
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2
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1
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1
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-
-
2
-
-
-
-
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-
-
1
1
-
-
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691390
Kawai
Structure and function of NAD ...
Arabidopsis thaliana, Saccharomyces cerevisiae
Biosci. Biotechnol. Biochem.
72
919-930
2008
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-
-
-
-
-
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1
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2
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2
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2
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1
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4
-
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2
-
-
2
-
-
693998
Panagiotou
Overexpression of a novel endo ...
Aspergillus nidulans, Aspergillus nidulans CBS 513.88 / FGSC A1513
Metab. Eng.
11
31-39
2008
-
-
1
-
-
-
-
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1
2
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4
-
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1
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4
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1
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4
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-
-
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706568
Panagiotou
-
Identification of NADH kinase ...
Fusarium oxysporum, Fusarium oxysporum F3
Process Biochem.
43
1114-1120
2008
-
-
-
-
-
-
8
2
-
1
2
-
-
6
-
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1
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1
-
4
1
1
-
1
2
1
1
1
1
-
1
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1
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8
-
2
-
1
2
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1
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1
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4
1
1
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1
2
1
1
1
1
-
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674616
Bieganowski
Synthetic lethal and biochemic ...
Saccharomyces cerevisiae
J. Biol. Chem.
281
22439-22445
2006
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1
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2
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1
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3
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1
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6
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1
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6
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676516
Chai
NADK3, a novel cytoplasmic sou ...
Arabidopsis thaliana
Plant J.
47
665-674
2006
-
-
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-
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1
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2
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5
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1
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1
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1
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5
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1
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657853
Turner
Identification, molecular clon ...
Arabidopsis thaliana
Biochem. J.
385
217-223
2005
-
-
1
-
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-
2
6
2
4
2
-
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5
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1
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3
3
-
8
1
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1
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1
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2
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6
2
4
2
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1
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3
3
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8
1
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1
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661696
Shi
Identification of ATP-NADH kin ...
Saccharomyces cerevisiae
FEBS J.
272
3337-3349
2005
-
-
-
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1
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3
-
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2
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1
1
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662376
Mori
Molecular conversion of NAD Ki ...
Micrococcus flavus, Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
280
24104-24112
2005
-
-
2
-
1
-
-
8
-
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161
-
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2
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4
-
6
1
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2
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2
2
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1
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8
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2
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4
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6
1
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676008
Berrin
Stress induces the expression ...
Arabidopsis thaliana
Mol. Genet. Genomics
273
10-19
2005
1
-
1
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2
2
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1
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1
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1
2
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2
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1
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1
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2
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1
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2
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2
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649040
Strand
POS5 gene of Saccharomyces cer ...
Saccharomyces cerevisiae
Eukaryot. Cell
2
809-820
2003
-
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1
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2
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1
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4
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