BRENDA - Enzyme Database

Polyphosphate:AMP phosphotransferase as a polyphosphate-dependent nucleoside monophosphate kinase in Acinetobacter johnsonii 210A

Shiba, T.; Itoh, H.; Kameda, A.; Kobayashi, K.; Kawazoe, Y.; Noguchi, T.; J. Bacteriol. 187, 1859-1865 (2005)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.4.B2
gene pap, construction of a cDNA library expressed in Escherichia coli, DNA and amino acid sequence determination and analysis, overexpression of pPAP2 from plasmid in Escherichia coli strain JM109
Acinetobacter johnsonii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.4.B2
ammonium sulfate
slightly inhibitory at 50 mM
Acinetobacter johnsonii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.B2
0.27
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
0.45
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
4.4
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.4.B2
Mg2+
specifically required, lowers the Km for the nucleotide substrate
Acinetobacter johnsonii
2.7.4.B2
additional information
other metal ions such as Mn2+, Fe2+, Ca2+, Cu2+, Zn2+, and Co2+ cannot enhance the activity
Acinetobacter johnsonii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.4.B2
55800
-
x * 55800, recombinant pPAP2, SDS-PAGE
Acinetobacter johnsonii
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.B2
AMP + [phosphate]n
Acinetobacter johnsonii
-
ADP + [phosphate]n-1
-
-
?
2.7.4.B2
AMP + [phosphate]n
Acinetobacter johnsonii 210A
-
ADP + [phosphate]n-1
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.4.B2
Acinetobacter johnsonii
Q83XD3
gene pap, plasmid containing the pap gene, designated pPAP2
-
2.7.4.B2
Acinetobacter johnsonii 210A
Q83XD3
gene pap, plasmid containing the pap gene, designated pPAP2
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.4.B2
recombinant pPAP2 dimer 22.1fold from Escherichia coli strain JM109 by ammonium sulfate fractionation, anion exchange and hydroxyapatite chromatography, and gel filtration
Acinetobacter johnsonii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.B2
2.48
-
crude extract of recombinant Escherichia coli strain JM109, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
24.9
-
purified recombinant PAP tetramer, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
54.9
-
purified recombinant PAP dimer, pH 8.0, 37°C
Acinetobacter johnsonii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.B2
AMP + [phosphate]n
-
722513
Acinetobacter johnsonii
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
best substrate
722513
Acinetobacter johnsonii
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
-
722513
Acinetobacter johnsonii 210A
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
best substrate
722513
Acinetobacter johnsonii 210A
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
CMP + [phosphate]n
0.09% of the activity with AMP
722513
Acinetobacter johnsonii
CDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
CMP + [phosphate]n
0.09% of the activity with AMP
722513
Acinetobacter johnsonii 210A
CDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dAMP + [phosphate]n
18% of the activity with AMP
722513
Acinetobacter johnsonii
dADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dCMP + [phosphate]n
0.008% of the activity with AMP
722513
Acinetobacter johnsonii
dCDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dGMP + [phosphate]n
2.6% of the activity with AMP
722513
Acinetobacter johnsonii
dGDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
GMP + [phosphate]n
10% of the activity with AMP
722513
Acinetobacter johnsonii
GDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
GMP + [phosphate]n
10% of the activity with AMP
722513
Acinetobacter johnsonii 210A
GDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
IMP + [phosphate]n
2.2% of the activity with AMP
722513
Acinetobacter johnsonii
IDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
additional information
PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively
722513
Acinetobacter johnsonii
?
-
-
-
-
2.7.4.B2
additional information
PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively
722513
Acinetobacter johnsonii 210A
?
-
-
-
-
2.7.4.B2
TMP + [phosphate]n
0.012% of the activity with AMP
722513
Acinetobacter johnsonii
TDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
UMP + [phosphate]n
0.13% of the activity with AMP
722513
Acinetobacter johnsonii
UDP + [phosphate]n-1
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.7.4.B2
?
x * 55800, recombinant pPAP2, SDS-PAGE
Acinetobacter johnsonii
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.4.B2
50
-
recombinant enzyme
Acinetobacter johnsonii
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
2.7.4.B2
30
50
the level of activity at 50°C is 3.2fold and 1.8fold higher than that at 30°C and 37°C, respectively
Acinetobacter johnsonii
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.4.B2
45
-
purified recombinant PAP, 10 min, 40% activity remaining without polyphosphate, 85% activity remaining in presence of 10 mM polyphosphate
Acinetobacter johnsonii
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.B2
35
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
151.7
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
166.7
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.4.B2
8
9
recombinant enzyme
Acinetobacter johnsonii
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
2.7.4.B2
7
11
purified recombinant enzyme, stable at
Acinetobacter johnsonii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.4.B2
gene pap, construction of a cDNA library expressed in Escherichia coli, DNA and amino acid sequence determination and analysis, overexpression of pPAP2 from plasmid in Escherichia coli strain JM109
Acinetobacter johnsonii
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.4.B2
ammonium sulfate
slightly inhibitory at 50 mM
Acinetobacter johnsonii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.B2
0.27
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
0.45
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
4.4
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.4.B2
Mg2+
specifically required, lowers the Km for the nucleotide substrate
Acinetobacter johnsonii
2.7.4.B2
additional information
other metal ions such as Mn2+, Fe2+, Ca2+, Cu2+, Zn2+, and Co2+ cannot enhance the activity
Acinetobacter johnsonii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.4.B2
55800
-
x * 55800, recombinant pPAP2, SDS-PAGE
Acinetobacter johnsonii
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.B2
AMP + [phosphate]n
Acinetobacter johnsonii
-
ADP + [phosphate]n-1
-
-
?
2.7.4.B2
AMP + [phosphate]n
Acinetobacter johnsonii 210A
-
ADP + [phosphate]n-1
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.4.B2
recombinant pPAP2 dimer 22.1fold from Escherichia coli strain JM109 by ammonium sulfate fractionation, anion exchange and hydroxyapatite chromatography, and gel filtration
Acinetobacter johnsonii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.B2
2.48
-
crude extract of recombinant Escherichia coli strain JM109, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
24.9
-
purified recombinant PAP tetramer, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
54.9
-
purified recombinant PAP dimer, pH 8.0, 37°C
Acinetobacter johnsonii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.B2
AMP + [phosphate]n
-
722513
Acinetobacter johnsonii
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
best substrate
722513
Acinetobacter johnsonii
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
-
722513
Acinetobacter johnsonii 210A
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
AMP + [phosphate]n
best substrate
722513
Acinetobacter johnsonii 210A
ADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
CMP + [phosphate]n
0.09% of the activity with AMP
722513
Acinetobacter johnsonii
CDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
CMP + [phosphate]n
0.09% of the activity with AMP
722513
Acinetobacter johnsonii 210A
CDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dAMP + [phosphate]n
18% of the activity with AMP
722513
Acinetobacter johnsonii
dADP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dCMP + [phosphate]n
0.008% of the activity with AMP
722513
Acinetobacter johnsonii
dCDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
dGMP + [phosphate]n
2.6% of the activity with AMP
722513
Acinetobacter johnsonii
dGDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
GMP + [phosphate]n
10% of the activity with AMP
722513
Acinetobacter johnsonii
GDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
GMP + [phosphate]n
10% of the activity with AMP
722513
Acinetobacter johnsonii 210A
GDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
IMP + [phosphate]n
2.2% of the activity with AMP
722513
Acinetobacter johnsonii
IDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
additional information
PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively
722513
Acinetobacter johnsonii
?
-
-
-
-
2.7.4.B2
additional information
PAP acts as a poly(P)-dependent NMP kinase, activity for AMP by PAP is about 10times greater than that for GMP and 770 and about 1100times greater than that for UMP and CMP. ATP production by the coupled reactions of recombinant PAP and purified Escherichia coli poly(P) kinases from AMP. The purified recombinant PAP enzyme has not only strong PAP activity but also shows poly(P)-dependent nucleoside monophosphate kinase activity, by which it converts ribonucleoside monophosphates and deoxyribonucleoside monophosphates into ribonucleoside diphosphates and deoxyribonucleoside diphosphates, respectively
722513
Acinetobacter johnsonii 210A
?
-
-
-
-
2.7.4.B2
TMP + [phosphate]n
0.012% of the activity with AMP
722513
Acinetobacter johnsonii
TDP + [phosphate]n-1
-
-
-
?
2.7.4.B2
UMP + [phosphate]n
0.13% of the activity with AMP
722513
Acinetobacter johnsonii
UDP + [phosphate]n-1
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.4.B2
?
x * 55800, recombinant pPAP2, SDS-PAGE
Acinetobacter johnsonii
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.4.B2
50
-
recombinant enzyme
Acinetobacter johnsonii
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
2.7.4.B2
30
50
the level of activity at 50°C is 3.2fold and 1.8fold higher than that at 30°C and 37°C, respectively
Acinetobacter johnsonii
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.4.B2
45
-
purified recombinant PAP, 10 min, 40% activity remaining without polyphosphate, 85% activity remaining in presence of 10 mM polyphosphate
Acinetobacter johnsonii
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.B2
35
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
151.7
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
166.7
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.4.B2
8
9
recombinant enzyme
Acinetobacter johnsonii
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
2.7.4.B2
7
11
purified recombinant enzyme, stable at
Acinetobacter johnsonii
General Information
EC Number
General Information
Commentary
Organism
2.7.4.B2
additional information
PAP contains two putative phosphate-binding motifs, P-loops. The similarity between PAP and the PPK2 homologues indicates that there is a common mechanism by which these enzymes use poly(P) for phosphorylation of nucleosides. The C-terminal region of PAP, including one P-loop (amino acids 286 to 292), is thought to be essential for poly(P)-dependent nucleotide phosphorylation. An additional P-loop in the N-terminal region (amino acids 45 to 51) of PAP may play a crucial role in binding with nucleoside monophosphate
Acinetobacter johnsonii
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.7.4.B2
additional information
PAP contains two putative phosphate-binding motifs, P-loops. The similarity between PAP and the PPK2 homologues indicates that there is a common mechanism by which these enzymes use poly(P) for phosphorylation of nucleosides. The C-terminal region of PAP, including one P-loop (amino acids 286 to 292), is thought to be essential for poly(P)-dependent nucleotide phosphorylation. An additional P-loop in the N-terminal region (amino acids 45 to 51) of PAP may play a crucial role in binding with nucleoside monophosphate
Acinetobacter johnsonii
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.4.B2
7.83
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
333.3
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
616.7
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.4.B2
7.83
-
GMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii
2.7.4.B2
333.3
-
AMP
recombinant PAP, pH 8.0, 37°C, in presence of 50 mM (NH4)2SO4
Acinetobacter johnsonii
2.7.4.B2
616.7
-
AMP
recombinant PAP, pH 8.0, 37°C
Acinetobacter johnsonii