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Literature summary for 2.7.1.86 extracted from

  • Ando, T.; Ohashi, K.; Ochiai, A.; Mikami, B.; Kawai, S.; Murata, K.
    Structural determinants of discrimination of NAD+ from NADH in yeast mitochondrial NADH kinase Pos5 (2011), J. Biol. Chem., 286, 29984-29992.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.1.86

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli RosettaBlue(DE3) cells Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) 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

Protein Variants

Protein Variants Comment 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 Comment 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 Comment Organism GeneOntology No. Textmining
mitochondrion
-
 Saccharomyces cerevisiae 5739
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46285
-
 2 * 46285, calculated from amino acid sequence Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae Q06892
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography and Superdex 200 gel filtration Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + NAD+ the enzyme has much higher NADH kinase than NAD kinase activity Saccharomyces cerevisiae ADP + NADP+
-
 ?
ATP + NADH the enzyme has much higher NADH kinase than NAD kinase activity Saccharomyces cerevisiae ADP + NADPH
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 46285, calculated from amino acid sequence Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
NADHK
-
 Saccharomyces cerevisiae
Pos5
-
 Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment 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 Comment 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