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Literature summary for 2.7.1.113 extracted from

  • Andersen, R.B.; Neuhard, J.
    Deoxynucleoside kinases encoded by the yaaG and yaaF genes of Bacillus subtilis. Substrate specificity and kinetic analysis of deoxyguanosine kinase with UTP as the preferred phosphate donor (2001), J. Biol. Chem., 276, 5518-5524.
    View publication on PubMed
Search for more literature for 2.7.1.113

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus subtilis
gene yaaG of the yaaG/yaaF operon, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli Bacillus subtilis

General Stability

General Stability Organism
bovine serum albumin stabilizes the enzyme in solution Bacillus subtilis
highly unstable at 0°C in diluted form, no loss of activity in the presence of bovine serum albumin Bacillus subtilis

Inhibitors

Inhibitors Comment Organism Structure
dGMP noncompetitive versus UTP and 2'-deoxyguanosine Bacillus subtilis
additional information no substrate inhibition by CTP, ATP, and GTP Bacillus subtilis
UDP competitive versus UTP, noncompetitive versus 2'-deoxyguanosine Bacillus subtilis
UTP substrate inhibition, overview Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0006
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis
0.0006
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with UTP as donor substrate Bacillus subtilis
0.0017
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with CTP as donor substrate Bacillus subtilis
0.006
-
 UTP pH 8.5, 37°C Bacillus subtilis
0.006
-
 UTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
0.0065
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate ATP Bacillus subtilis
0.0065
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with ATP as donor substrate Bacillus subtilis
0.01
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate GTP Bacillus subtilis
0.0104
-
 2'-deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with GTP as donor substrate Bacillus subtilis
0.017
-
 2'-deoxyguanosine pH 8.5, 37°C, cosubstrate CTP Bacillus subtilis
0.035
-
 CTP pH 8.5, 37°C Bacillus subtilis
0.035
-
 CTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
0.036
-
 ATP pH 8.5, 37°C Bacillus subtilis
0.036
-
 ATP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
0.046
-
 GTP pH 8.5, 37°C Bacillus subtilis
0.046
-
 GTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for activity Bacillus subtilis
Mg2+ MgNTP2- is most probably the true phosphate donor Bacillus subtilis
Mg2+ required for activity, no inhibition by excess Mg2+, at 5 mM Bacillus subtilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
24147
-
 2 * 24147, electrospray mass spectrometry Bacillus subtilis
24147
-
 2 * 24147, recombinant enzyme, electrospray mass spectrometry Bacillus subtilis
49000
-
 recombinant enzyme, gel filtration Bacillus subtilis
49000
-
 recombinant deoxyguanosine kinase, gel filtration Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-
Bacillus subtilis
-
 gene yaaG
-

Purification (Commentary)

Purification (Comment) Organism
recombinant deoxyguanosine kinase, BlueA dye affinity, phenyl-Sepharose Bacillus subtilis
recombinant enzyme from Escherichia coli by affinity and phenyl resin chromatography to homogeneity Bacillus subtilis

Reaction

Reaction Comment Organism Reaction ID
ATP + deoxyguanosine = ADP + dGMP sequential Bi Bi mechanism Bacillus subtilis
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.54
-
 enzyme in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyguanosine Bacillus subtilis
2.3
-
 dAdo/dCyd kinase in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyadenosine or 2'-deoxycytidine Bacillus subtilis
8.1
-
 recombinant deoxyguanosine kinase Bacillus subtilis
8.1
-
 purified recombinant enzyme, substrate 2'-deoxyguanosine Bacillus subtilis

Storage Stability

Storage Stability Organism
-20°C, more than 1 mg/ml protein, 50% glycerol, 1 year, no loss of activity Bacillus subtilis
-20°C, purified recombinant enzyme at 1 mg/ml, Tris-HCl, pH 7.8, 50% v/v glycerol, at least 1 year without loss of activity Bacillus subtilis
0°C, the diluted enzyme is highly unstable, but in presence of 1 mg/ml bovine serum albumin, the diluted enzyme is stable for months Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 2'-deoxyguanosine
-
 Bacillus subtilis ADP + 2'-deoxyguanosine 5'-phosphate
-
 ?
ATP + deoxyadenosine 3% of activity with deoxyguanosine Bacillus subtilis ADP + dAMP
-
 ?
ATP + deoxycytidine recombinant deoxyguanosine kinase, 3% of activity with deoxyguanosine Bacillus subtilis ADP + dCMP
-
 ?
ATP + deoxyinosine 63% of activity with deoxyguanosine Bacillus subtilis ADP + dIMP
-
 ?
ATP + guanosine recombinant deoxyguanosine kinase, 7% of activity with deoxyguanosine Bacillus subtilis ADP + GMP
-
 ?
CTP + 2'-deoxyguanosine
-
 Bacillus subtilis CDP + 2'-deoxyguanosine 5'-phosphate
-
 ?
GTP + 2'-deoxyadenosine very low activity Bacillus subtilis GDP + 2'-deoxyadenosine 5'-phosphate
-
 ?
GTP + 2'-deoxycytidine very low activity Bacillus subtilis GDP + 2'-deoxycytidine 5'-phosphate
-
 ?
GTP + 2'-deoxyguanosine
-
 Bacillus subtilis GDP + 2'-deoxyguanosine 5'-phosphate
-
 ?
GTP + 2'-deoxyinosine
-
 Bacillus subtilis GDP + 2'-deoxyinosine 5'-phosphate
-
 ?
GTP + guanosine very low activity Bacillus subtilis GDP + guanosine 5'-phosphate
-
 ?
additional information substrate specificity, the enzyme utilizes all naturally occurring ribo-and deoxyribonucleosides triphosphates, with the exception of dGTP, as phosphate donors, and shows poor activity with thymidine, inosine, adenosine, 2,3-dideoxyguanosine, 2,3-dideoxyinosine, 2,3-dideoxyadenosine, and hypoxanthine arabinoside as acceptor substrates Bacillus subtilis ?
-
 ?
UTP + 2'-deoxyguanosine preferred substrates Bacillus subtilis UDP + 2'-deoxyguanosine 5'-phosphate
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 24147, electrospray mass spectrometry Bacillus subtilis
dimer 2 * 24147, recombinant enzyme, electrospray mass spectrometry Bacillus subtilis

Synonyms

Synonyms Comment Organism
deoxyguanosine kinase
-
 Bacillus subtilis
DGK
-
 Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.41
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis
1.41
-
 UTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
1.41
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with UTP as donor substrate Bacillus subtilis
1.53
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate ATP Bacillus subtilis
1.53
-
 ATP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
1.53
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with ATP as donor substrate Bacillus subtilis
1.69
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate GTP Bacillus subtilis
1.69
-
 GTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
1.69
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with GTP as donor substrate Bacillus subtilis
2.25
-
 deoxyguanosine pH 8.5, 37°C, cosubstrate CTP Bacillus subtilis
2.25
-
 CTP pH 8.5, 37°C, recombinant enzyme, with 2'-deoxyguanosine as acceptor substrate Bacillus subtilis
2.25
-
 2'deoxyguanosine pH 8.5, 37°C, recombinant enzyme, with CTP as donor substrate Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Bacillus subtilis
9
-
 broad optimum Bacillus subtilis

pH Range

pH Minimum pH Maximum Comment Organism
5.6 9 more than 80% of maximal activity at pH 7.5 and pH 11.5, respectively, 60% of maximal activity at pH 6.0, complete loss of activity below pH 5.6 Bacillus subtilis
6 11.5 over 80% of maximal activity at pH 7.5 and pH 11.5, about 60% of maximal activity at pH 6.0, enzyme is completely inactive below pH 5.6 Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Bacillus subtilis
CTP
-
 Bacillus subtilis
GTP
-
 Bacillus subtilis
additional information the enzyme utilizes all naturally occurring ribo-and deoxyribonucleosides triphosphates, with the exception of dGTP, as phosphate donors Bacillus subtilis
UTP
-
 Bacillus subtilis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
 additional information product inhibition of the enzyme, overview Bacillus subtilis
0.063
-
 UDP pH 8.5, 37°C, cosubstrate UTP Bacillus subtilis