BRENDA - Enzyme Database

Purification and molecular characterization of a novel diadenosine 5',5'''-P1,P4-tetraphosphate phosphorylase from Mycobacterium tuberculosis H37Rv

Mori, S.; Shibayama, K.; Wachino, J.; Arakawa, Y.; Protein Expr. Purif. 69, 99-105 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.7.53
expressed in Escherichia coli BL21(DE3)pLysS cells
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.1
-
P1,P4-bis(5'-adenosyl) tetraphosphate
in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
2.7.7.53
0.94
-
phosphate
in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.7.53
Ca2+
Rv2613c requires bivalent metal ions such as Ca2+ to exert its Ap4A phosphorylase activity (64% activity in the presence of 2 mM Ca2+)
Mycobacterium tuberculosis
2.7.7.53
Co2+
Rv2613c requires bivalent metal ions such as Co2+ to exert its Ap4A phosphorylase activity (68% activity in the presence of 2 mM Co2+)
Mycobacterium tuberculosis
2.7.7.53
Mg2+
Rv2613c requires bivalent metal ions such as Mg2+ to exert its Ap4A phosphorylase activity (34% activity in the presence of 2 mM Mg2+)
Mycobacterium tuberculosis
2.7.7.53
Mn2+
Rv2613c requires bivalent metal ions such as Mn2+ to exert its Ap4A phosphorylase activity (100% activity in the presence of 2 mM Mn2+)
Mycobacterium tuberculosis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.7.53
24683
-
4 * 24683, MALDI-TOF mass spectrometry
Mycobacterium tuberculosis
2.7.7.53
100000
-
gel filtration
Mycobacterium tuberculosis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.7.53
Mycobacterium tuberculosis
P9WMK9
-
-
2.7.7.53
Mycobacterium tuberculosis H37Rv
P9WMK9
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.7.53
HisTrap HP column chromatography and Sephacryl S-200 gel filtration
Mycobacterium tuberculosis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.7.53
3.01
-
crude enzyme, using P1,P4-bis(5'-adenosyl) tetraphosphate as substrate, in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
2.7.7.53
10.84
-
after 3.6fold purification, using P1,P4-bis(5'-adenosyl) tetraphosphate as substrate, in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.7.53
additional information
the enzyme shows no hydrolysis of AMP-NH2, and almost no activity with adenosine tetraphosphate, ATP, ADP-ribose, ADP-glucose, guanosine tetraphosphate, guanosine 5'-triphosphate, GDP-glucose, GDP-mannose, adenylyl imidodiphosphate, and guanylyl imidodiphosphate
713455
Mycobacterium tuberculosis
?
-
-
-
-
2.7.7.53
additional information
the enzyme shows no hydrolysis of AMP-NH2, and almost no activity with adenosine tetraphosphate, ATP, ADP-ribose, ADP-glucose, guanosine tetraphosphate, guanosine 5'-triphosphate, GDP-glucose, GDP-mannose, adenylyl imidodiphosphate, and guanylyl imidodiphosphate
713455
Mycobacterium tuberculosis H37Rv
?
-
-
-
-
2.7.7.53
P1,P3-bis(5'-adenosyl) triphosphate + phosphate
19% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
ADP
-
-
-
?
2.7.7.53
P1,P3-bis(5'-adenosyl) triphosphate + phosphate
19% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
ADP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
100% activity
713455
Mycobacterium tuberculosis
ATP + ADP
-
-
-
ir
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
100% activity
713455
Mycobacterium tuberculosis H37Rv
ATP + ADP
-
-
-
ir
2.7.7.53
P1,P4-bis(5'-guanosyl) tetraphosphate + phosphate
107% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
GTP + GDP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-guanosyl) tetraphosphate + phosphate
107% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
GTP + GDP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-adenosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine tetraphosphate + ADP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-adenosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
adenosine tetraphosphate + ADP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-guanosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
guanosine tetraphosphate + GDP
-
-
-
?
2.7.7.53
P1,P6-bis(5'-adenosyl) hexaphosphate + phosphate
25% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine pentaphosphate + ADP
-
-
-
?
2.7.7.53
P1-(5'-adenosyl)P4-(5'-guanosyl) tetraphosphate + phosphate
100% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
ATP + GDP
-
-
-
?
2.7.7.53
P1-(5'-adenosyl)P5-(5'-guanosyl) pentaphosphate + phosphate
76% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine tetraphosphate + GDP
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.7.7.53
homotetramer
4 * 24683, MALDI-TOF mass spectrometry
Mycobacterium tuberculosis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.7.53
30
-
-
Mycobacterium tuberculosis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.7.53
65
-
the activity completely disappears after treatment at 65°C for 10 min
Mycobacterium tuberculosis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.7.53
8
-
-
Mycobacterium tuberculosis
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.7.53
7.4
10
-
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.7.53
expressed in Escherichia coli BL21(DE3)pLysS cells
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.1
-
P1,P4-bis(5'-adenosyl) tetraphosphate
in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
2.7.7.53
0.94
-
phosphate
in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.7.53
Ca2+
Rv2613c requires bivalent metal ions such as Ca2+ to exert its Ap4A phosphorylase activity (64% activity in the presence of 2 mM Ca2+)
Mycobacterium tuberculosis
2.7.7.53
Co2+
Rv2613c requires bivalent metal ions such as Co2+ to exert its Ap4A phosphorylase activity (68% activity in the presence of 2 mM Co2+)
Mycobacterium tuberculosis
2.7.7.53
Mg2+
Rv2613c requires bivalent metal ions such as Mg2+ to exert its Ap4A phosphorylase activity (34% activity in the presence of 2 mM Mg2+)
Mycobacterium tuberculosis
2.7.7.53
Mn2+
Rv2613c requires bivalent metal ions such as Mn2+ to exert its Ap4A phosphorylase activity (100% activity in the presence of 2 mM Mn2+)
Mycobacterium tuberculosis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.7.53
24683
-
4 * 24683, MALDI-TOF mass spectrometry
Mycobacterium tuberculosis
2.7.7.53
100000
-
gel filtration
Mycobacterium tuberculosis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.7.53
HisTrap HP column chromatography and Sephacryl S-200 gel filtration
Mycobacterium tuberculosis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.7.53
3.01
-
crude enzyme, using P1,P4-bis(5'-adenosyl) tetraphosphate as substrate, in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
2.7.7.53
10.84
-
after 3.6fold purification, using P1,P4-bis(5'-adenosyl) tetraphosphate as substrate, in 50 mM HEPES-Na (pH 7.6), 1 mM MnCl2, at 37°C
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.7.53
additional information
the enzyme shows no hydrolysis of AMP-NH2, and almost no activity with adenosine tetraphosphate, ATP, ADP-ribose, ADP-glucose, guanosine tetraphosphate, guanosine 5'-triphosphate, GDP-glucose, GDP-mannose, adenylyl imidodiphosphate, and guanylyl imidodiphosphate
713455
Mycobacterium tuberculosis
?
-
-
-
-
2.7.7.53
additional information
the enzyme shows no hydrolysis of AMP-NH2, and almost no activity with adenosine tetraphosphate, ATP, ADP-ribose, ADP-glucose, guanosine tetraphosphate, guanosine 5'-triphosphate, GDP-glucose, GDP-mannose, adenylyl imidodiphosphate, and guanylyl imidodiphosphate
713455
Mycobacterium tuberculosis H37Rv
?
-
-
-
-
2.7.7.53
P1,P3-bis(5'-adenosyl) triphosphate + phosphate
19% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
ADP
-
-
-
?
2.7.7.53
P1,P3-bis(5'-adenosyl) triphosphate + phosphate
19% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
ADP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
100% activity
713455
Mycobacterium tuberculosis
ATP + ADP
-
-
-
ir
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
100% activity
713455
Mycobacterium tuberculosis H37Rv
ATP + ADP
-
-
-
ir
2.7.7.53
P1,P4-bis(5'-guanosyl) tetraphosphate + phosphate
107% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
GTP + GDP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-guanosyl) tetraphosphate + phosphate
107% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
GTP + GDP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-adenosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine tetraphosphate + ADP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-adenosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis H37Rv
adenosine tetraphosphate + ADP
-
-
-
?
2.7.7.53
P1,P5-bis(5'-guanosyl) pentaphosphate + phosphate
106% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
guanosine tetraphosphate + GDP
-
-
-
?
2.7.7.53
P1,P6-bis(5'-adenosyl) hexaphosphate + phosphate
25% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine pentaphosphate + ADP
-
-
-
?
2.7.7.53
P1-(5'-adenosyl)P4-(5'-guanosyl) tetraphosphate + phosphate
100% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
ATP + GDP
-
-
-
?
2.7.7.53
P1-(5'-adenosyl)P5-(5'-guanosyl) pentaphosphate + phosphate
76% activity compared to P1,P4-bis(5'-adenosyl) tetraphosphate
713455
Mycobacterium tuberculosis
adenosine tetraphosphate + GDP
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.7.53
homotetramer
4 * 24683, MALDI-TOF mass spectrometry
Mycobacterium tuberculosis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.7.53
30
-
-
Mycobacterium tuberculosis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.7.53
65
-
the activity completely disappears after treatment at 65°C for 10 min
Mycobacterium tuberculosis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.7.53
8
-
-
Mycobacterium tuberculosis
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.7.53
7.4
10
-
Mycobacterium tuberculosis