BRENDA - Enzyme Database
show all sequences of 2.7.1.B26

Mechanism of aminoglycoside 3'-phosphotransferase type IIIa: His188 is not a phosphate-accepting residue

Thompson, P.R.; Hughes, D.W.; Wright, G.R.; Chem. Biol. 3, 747-755 (1996)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Enterococcus faecium
Engineering
Amino acid exchange
Commentary
Organism
H123A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H188A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H78A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H82A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
Inhibitors
Inhibitors
Commentary
Organism
Structure
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide
10 mM, 2% residual activity
Enterococcus faecium
4-chloromercuribenzoic acid
0.5 mM, 3% residual activity
Enterococcus faecium
diethyldicarbonate
2.5 mM, 10% residual activity
Enterococcus faecium
kanamycin A
-
Enterococcus faecium
N-ethylmaleimide
0.5 mM, 47% residual activity
Enterococcus faecium
neomycin B
-
Enterococcus faecium
Phenylglyoxal
2.5 mM, 10% residual activity
Enterococcus faecium
Tetranitromethane
2.5 mM, 23% residual activity
Enterococcus faecium
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00029
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.00092
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.00116
-
ATP
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.0013
-
neomycin B
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0024
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0032
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.0051
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.00772
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.011
-
ATP
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.012
-
ATP
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0126
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.0174
-
kanamycin A
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.027
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.0277
-
ATP
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.0335
-
ATP
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.093
-
amikacin
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.11
-
amikacin
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.184
-
amikacin
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.206
-
amikacin
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.245
-
amikacin
wild-type, pH 7.5, 37°C
Enterococcus faecium
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Enterococcus faecium
E9LMR8
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + amikacin
-
733646
Enterococcus faecium
ADP + amikacin 3'-phosphate
-
-
-
?
ATP + kanamycin A
-
733646
Enterococcus faecium
ADP + kanamycin A 3'-phosphate
-
-
-
?
ATP + neomycin B
-
733646
Enterococcus faecium
ADP + neomycin B 3'-phosphate
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.35
-
ATP
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.35
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.49
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.5
-
amikacin
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.57
-
ATP
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.66
-
ATP
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.67
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.69
-
neomycin B
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.83
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.87
-
amikacin
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
1.2
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
1.74
-
amikacin
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
1.76
-
ATP
wild-type, pH 7.5, 37°C
Enterococcus faecium
1.79
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.04
-
amikacin
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
2.08
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.46
-
amikacin
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.46
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
3.36
-
kanamycin A
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
3.75
-
ATP
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.68
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
1.84
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
2.65
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.74
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
5.03
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
6.38
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
6.7
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
6.95
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Enterococcus faecium
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
H123A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H188A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H78A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
H82A
mutation has minimal effects on phosphorylation of several aminoglycosides
Enterococcus faecium
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide
10 mM, 2% residual activity
Enterococcus faecium
4-chloromercuribenzoic acid
0.5 mM, 3% residual activity
Enterococcus faecium
diethyldicarbonate
2.5 mM, 10% residual activity
Enterococcus faecium
kanamycin A
-
Enterococcus faecium
N-ethylmaleimide
0.5 mM, 47% residual activity
Enterococcus faecium
neomycin B
-
Enterococcus faecium
Phenylglyoxal
2.5 mM, 10% residual activity
Enterococcus faecium
Tetranitromethane
2.5 mM, 23% residual activity
Enterococcus faecium
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.68
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
1.84
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
2.65
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.74
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
5.03
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
6.38
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
6.7
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
6.95
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00029
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.00092
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.00116
-
ATP
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.0013
-
neomycin B
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0024
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0032
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.0051
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.00772
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.011
-
ATP
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.012
-
ATP
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.0126
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.0174
-
kanamycin A
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.027
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.0277
-
ATP
wild-type, pH 7.5, 37°C
Enterococcus faecium
0.0335
-
ATP
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.093
-
amikacin
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.11
-
amikacin
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.184
-
amikacin
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
0.206
-
amikacin
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.245
-
amikacin
wild-type, pH 7.5, 37°C
Enterococcus faecium
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + amikacin
-
733646
Enterococcus faecium
ADP + amikacin 3'-phosphate
-
-
-
?
ATP + kanamycin A
-
733646
Enterococcus faecium
ADP + kanamycin A 3'-phosphate
-
-
-
?
ATP + neomycin B
-
733646
Enterococcus faecium
ADP + neomycin B 3'-phosphate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.35
-
ATP
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.35
-
kanamycin A
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.49
-
kanamycin A
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.5
-
amikacin
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.57
-
ATP
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.66
-
ATP
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.67
-
kanamycin A
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
0.69
-
neomycin B
mutant H82A, pH 7.5, 37°C
Enterococcus faecium
0.83
-
neomycin B
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
0.87
-
amikacin
mutant H123A, pH 7.5, 37°C
Enterococcus faecium
1.2
-
neomycin B
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
1.74
-
amikacin
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
1.76
-
ATP
wild-type, pH 7.5, 37°C
Enterococcus faecium
1.79
-
kanamycin A
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.04
-
amikacin
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
2.08
-
neomycin B
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.46
-
amikacin
wild-type, pH 7.5, 37°C
Enterococcus faecium
2.46
-
neomycin B
mutant H78A, pH 7.5, 37°C
Enterococcus faecium
3.36
-
kanamycin A
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
3.75
-
ATP
mutant H188A, pH 7.5, 37°C
Enterococcus faecium
Other publictions for EC 2.7.1.B26
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733482
Shao
Characterization of a key amin ...
Micromonospora echinospora, Micromonospora echinospora ATCC 15835
Bioorg. Med. Chem. Lett.
23
1438-1441
2013
-
-
1
-
-
-
-
4
-
-
1
-
-
2
-
-
1
-
-
-
-
-
10
1
-
-
-
4
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
1
-
-
-
-
10
1
-
-
-
4
-
-
-
-
-
-
-
-
4
4
733103
McKay
Recognition of aminoglycoside ...
Enterococcus faecium
Antimicrob. Agents Chemother.
40
2648-2650
1996
-
-
-
-
-
-
2
6
-
-
-
-
-
1
-
1
-
-
-
-
-
-
7
-
-
-
-
6
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
2
2
6
-
-
-
-
-
-
1
-
-
-
-
-
7
-
-
-
-
6
-
-
-
-
-
-
-
-
5
5
733646
Thompson
Mechanism of aminoglycoside 3' ...
Enterococcus faecium
Chem. Biol.
3
747-755
1996
-
-
1
-
4
-
8
20
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
20
-
-
-
-
8
-
-
-
-
1
-
-
4
-
-
8
8
20
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
20
-
-
-
-
-
-
-
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-
-