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Literature summary for 2.7.1.23 extracted from

  • Jeelani, G.; Husain, A.; Sato, D.; Soga, T.; Suematsu, M.; Nozaki, T.
    Biochemical and functional characterization of novel NADH kinase in the enteric protozoan parasite Entamoeba histolytica (2013), Biochimie, 95, 309-319.
    View publication on PubMed
Search for more literature for 2.7.1.23

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Entamoeba histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
 NADH in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
0.4
-
 ATP with NADH as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
1.59
-
 NAD+ in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
1.74
-
 ATP with NAD+ as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ 43% activity at 5 mM compared to Mg2+ Entamoeba histolytica
Co2+ 36% activity at 5 mM compared to Mg2+ Entamoeba histolytica
Cu2+ 2% activity at 5 mM compared to Mg2+ Entamoeba histolytica
Fe2+ 83% activity at 5 mM compared to Mg2+ Entamoeba histolytica
Mg2+ preferred cation, 100% activity at 5 mM Entamoeba histolytica
Mn2+ 15% activity at 5 mM compared to Mg2+ Entamoeba histolytica
additional information no activity is detected in presence of Li+, Na+, and K+ Entamoeba histolytica
Ni2+ 6% activity at 5 mM compared to Mg2+ Entamoeba histolytica
Zn2+ 10% activity at 5 mM compared to Mg2+ Entamoeba histolytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29500
-
 x * 29500, calculated from amino acid sequence Entamoeba histolytica
32000
-
 x * 32000, recombinant enzyme, SDS-PAGE Entamoeba histolytica

Organism

Organism UniProt Comment Textmining
Entamoeba histolytica C4LSN1
-
-
Entamoeba histolytica HM1: IMSS cl 6 C4LSN1
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography, gel filtration Entamoeba histolytica

Storage Stability

Storage Stability Organism
-30 or -80°C, 10-15% (v/v) glycerol, at least 3 months, no loss of activity Entamoeba histolytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + NADH 13% activity compared to ATP Entamoeba histolytica AMP + NADPH
-
 ?
ATP + NAD+ 50% activity compared to NADH Entamoeba histolytica ADP + NADP+
-
 ?
ATP + NAD+ 50% activity compared to NADH Entamoeba histolytica HM1: IMSS cl 6 ADP + NADP+
-
 ?
ATP + NADH 100% activity Entamoeba histolytica ADP + NADPH
-
 ?
ATP + NADH 100% activity Entamoeba histolytica HM1: IMSS cl 6 ADP + NADPH
-
 ?
CTP + NADH 74% activity compared to ATP Entamoeba histolytica CDP + NADPH
-
 ?
CTP + NADH 74% activity compared to ATP Entamoeba histolytica HM1: IMSS cl 6 CDP + NADPH
-
 ?
dATP + NADH 103% activity compared to ATP Entamoeba histolytica dADP + NADPH
-
 ?
GTP + NADH 102% activity compared to ATP Entamoeba histolytica GDP + NADPH
-
 ?
additional information the enzyme does not use AMP, CMP or inorganic polyphosphates (diphosphate, tripolyphosphate, trimetaphosphate, hexametaphosphate, metaphosphate, and polyphosphate) as the phosphoryl donor. Glucose 6-phosphate and phosphoenolpyruvate are also inert as phosphoryl donors Entamoeba histolytica ?
-
 ?
additional information the enzyme does not use AMP, CMP or inorganic polyphosphates (diphosphate, tripolyphosphate, trimetaphosphate, hexametaphosphate, metaphosphate, and polyphosphate) as the phosphoryl donor. Glucose 6-phosphate and phosphoenolpyruvate are also inert as phosphoryl donors Entamoeba histolytica HM1: IMSS cl 6 ?
-
 ?
TTP + NADH 50% activity compared to ATP Entamoeba histolytica TDP + NADPH
-
 ?
UTP + NADH 69% activity compared to ATP Entamoeba histolytica UDP + NADPH
-
 ?
UTP + NADH 69% activity compared to ATP Entamoeba histolytica HM1: IMSS cl 6 UDP + NADPH
-
 ?

Subunits

Subunits Comment Organism
? x * 32000, recombinant enzyme, SDS-PAGE Entamoeba histolytica
? x * 29500, calculated from amino acid sequence Entamoeba histolytica

Synonyms

Synonyms Comment Organism
NAD(H) kinase
-
 Entamoeba histolytica
NADHK
-
 Entamoeba histolytica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.9
-
 NAD+ in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
1.9
-
 NADH in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
2.02
-
 ATP with NAD+ as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
2.68
-
 ATP with NADH as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Entamoeba histolytica

pI Value

Organism Comment pI Value Maximum pI Value
Entamoeba histolytica calculated from amino acid sequence
-
 6.5

General Information

General Information Comment Organism
physiological function enzyme overexpression represses the production of intracellular reactive oxygen specie upon H2O2 exposure by 40% Entamoeba histolytica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.16
-
 ATP with NAD+ as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
1.2
-
 NAD+ in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
6.72
-
 ATP with NADH as cosubstrate, in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica
56.48
-
 NADH in 100 mM Tris-HCl (pH 7.5), at 37°C Entamoeba histolytica