BRENDA - Enzyme Database

Structural insights into the novel diadenosine 5',5'''-P1,P4-tetraphosphate phosphorylase from Mycobacterium tuberculosis H37Rv

Mori, S.; Shibayama, K.; Wachino, J.; Arakawa, Y.; J. Mol. Biol. 410, 93-104 (2011)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.7.53
expression in Escherichia coli
Mycobacterium tuberculosis
Crystallization (Commentary)
EC Number
Crystallization
Organism
2.7.7.53
crystal structure at 1.89 A resolution by using single-wavelength anomalous dispersion. Asn139, Gly146, and Ser147 are important active-site residues, Asn139 has a critical role in catalysis. The position of Gly146 might influence the phosphorylase activity. The tetrameric structure and the presence of Trp160 might be essential for the formation of the Ap4A binding site
Mycobacterium tuberculosis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.7.53
G146Q
about 20% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
G146S
about 30% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
N139A
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
N139Q
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
N139Q/S147T
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
S147A
about 8% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
S147T
about 15% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
W160A
complete loss of activity
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.07
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.08
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C; mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.1
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.11
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 37°C
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
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
-
722980
Mycobacterium tuberculosis
ADP + ATP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
-
722980
Mycobacterium tuberculosis H37Rv
ADP + ATP
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.48
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.38
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.49
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.99
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
8.48
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.7.53
expression in Escherichia coli
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.7.7.53
crystal structure at 1.89 A resolution by using single-wavelength anomalous dispersion. Asn139, Gly146, and Ser147 are important active-site residues, Asn139 has a critical role in catalysis. The position of Gly146 might influence the phosphorylase activity. The tetrameric structure and the presence of Trp160 might be essential for the formation of the Ap4A binding site
Mycobacterium tuberculosis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.7.53
G146Q
about 20% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
G146S
about 30% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
N139A
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
N139Q
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
N139Q/S147T
complete loss of activity
Mycobacterium tuberculosis
2.7.7.53
S147A
about 8% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
S147T
about 15% of wild-type activity
Mycobacterium tuberculosis
2.7.7.53
W160A
complete loss of activity
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.07
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.08
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C; mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.1
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
0.11
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 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
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
-
722980
Mycobacterium tuberculosis
ADP + ATP
-
-
-
?
2.7.7.53
P1,P4-bis(5'-adenosyl) tetraphosphate + phosphate
-
722980
Mycobacterium tuberculosis H37Rv
ADP + ATP
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.7.53
0.48
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.38
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.49
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
1.99
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
8.48
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.7.53
6.9
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
13.6
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
17.25
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
24.88
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
84.8
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis
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
2.7.7.53
6.9
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147A, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
13.6
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant S147T, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
17.25
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146Q, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
24.88
-
P1,P4-bis(5'-adenosyl) tetraphosphate
mutant G146S, pH 7.6, 37°C
Mycobacterium tuberculosis
2.7.7.53
84.8
-
P1,P4-bis(5'-adenosyl) tetraphosphate
wild-type, pH 7.6, 37°C
Mycobacterium tuberculosis