BRENDA - Enzyme Database
show all sequences of 2.7.1.162

Open-close structural change upon ligand binding and two magnesium ions required for the catalysis of N-acetylhexosamine 1-kinase

Sato, M.; Arakawa, T.; Nam, Y.W.; Nishimoto, M.; Kitaoka, M.; Fushinobu, S.; Biochim. Biophys. Acta 1854, 333-340 (2015)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene lnpB, recombinant expression of C-terminally His6-tagged NahK residues 1-359 in Escherichia coli strain BL21 CodonPlus (DE3)-RIL, expression of the (SeMet)-labeled protein in Escherichia coli strain B834 (DE3)
Bifidobacterium longum
Crystallization (Commentary)
Crystallization
Organism
purified recombinant wild-type enzyme in apoform, or SeMet-labeled NahK complexed with ATP, sitting drop vapor diffusion method, mixing of 500 nl protein solution containing 25 mg/mL NahK and with or without 10 mM ATP with an equal volume of reservoir solution containing 0.2 M magnesium acetate, 0.1 M sodium cacodylate, pH 6.5, and 20% PEG 8000, for ADP complex crystals, 10 mM ADP is used instead of ATP, 20°C, X-ray diffraction structure determination and analysis at 1.80-2.05 A resolution
Bifidobacterium longum
Engineering
Amino acid exchange
Commentary
Organism
D208A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
D228A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
K210A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
N213A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.005
-
ATP
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.038
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
0.1
-
ATP
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
0.35
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.9
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
1.8
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
2.5
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
3.14
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
requirement of the second magnesium ion, the enzyme has two magnesium binding sites, Mg1 and Mg2, structure overview
Bifidobacterium longum
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + N-acetyl-D-hexosamine
Bifidobacterium longum
-
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
?
ATP + N-acetyl-D-hexosamine
Bifidobacterium longum JCM 1217
-
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Bifidobacterium longum
E8MF12
gene lnpB
-
Bifidobacterium longum JCM 1217
E8MF12
gene lnpB
-
Purification (Commentary)
Commentary
Organism
recombinant C-terminally His6-tagged wild-type and SeMet-labeled NahK residues 1-359 enzymes from Escherichia coli by nickel affinity chromatography and gel filtration
Bifidobacterium longum
Reaction
Reaction
Commentary
Organism
ATP + N-acetyl-D-hexosamine = ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
open–close conformational change at the active site, structure overview
Bifidobacterium longum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + N-acetyl-D-glucosamine
-
737770
Bifidobacterium longum
ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-glucosamine
-
737770
Bifidobacterium longum JCM 1217
ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-hexosamine
-
737770
Bifidobacterium longum
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-hexosamine
-
737770
Bifidobacterium longum JCM 1217
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
-
?
Subunits
Subunits
Commentary
Organism
dimer
-
Bifidobacterium longum
More
enzyme domain architecture, overview
Bifidobacterium longum
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Bifidobacterium longum
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.11
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.12
-
ATP
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
5.9
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
6.4
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
8.8
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
9.9
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
13.6
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
16.3
-
ATP
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Bifidobacterium longum
Cloned(Commentary) (protein specific)
Commentary
Organism
gene lnpB, recombinant expression of C-terminally His6-tagged NahK residues 1-359 in Escherichia coli strain BL21 CodonPlus (DE3)-RIL, expression of the (SeMet)-labeled protein in Escherichia coli strain B834 (DE3)
Bifidobacterium longum
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant wild-type enzyme in apoform, or SeMet-labeled NahK complexed with ATP, sitting drop vapor diffusion method, mixing of 500 nl protein solution containing 25 mg/mL NahK and with or without 10 mM ATP with an equal volume of reservoir solution containing 0.2 M magnesium acetate, 0.1 M sodium cacodylate, pH 6.5, and 20% PEG 8000, for ADP complex crystals, 10 mM ADP is used instead of ATP, 20°C, X-ray diffraction structure determination and analysis at 1.80-2.05 A resolution
Bifidobacterium longum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D208A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
D228A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
K210A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
N213A
site-directed mutagenesis, the mutant shows altered activity and kinetics compared to the wild-type enzyme
Bifidobacterium longum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.005
-
ATP
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.038
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
0.1
-
ATP
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
0.35
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.9
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
1.8
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
2.5
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
3.14
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
requirement of the second magnesium ion, the enzyme has two magnesium binding sites, Mg1 and Mg2, structure overview
Bifidobacterium longum
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + N-acetyl-D-hexosamine
Bifidobacterium longum
-
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
?
ATP + N-acetyl-D-hexosamine
Bifidobacterium longum JCM 1217
-
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant C-terminally His6-tagged wild-type and SeMet-labeled NahK residues 1-359 enzymes from Escherichia coli by nickel affinity chromatography and gel filtration
Bifidobacterium longum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + N-acetyl-D-glucosamine
-
737770
Bifidobacterium longum
ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-glucosamine
-
737770
Bifidobacterium longum JCM 1217
ADP + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-hexosamine
-
737770
Bifidobacterium longum
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
-
?
ATP + N-acetyl-D-hexosamine
-
737770
Bifidobacterium longum JCM 1217
ADP + N-acetyl-alpha-D-hexosamine 1-phosphate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
-
Bifidobacterium longum
More
enzyme domain architecture, overview
Bifidobacterium longum
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Bifidobacterium longum
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.11
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
0.12
-
ATP
pH 8.0, 37°C, recombinant mutant K210A, at 5.0 mM MgCl2
Bifidobacterium longum
5.9
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
6.4
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 5.0 mM MgCl2
Bifidobacterium longum
8.8
-
ATP
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
9.9
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
13.6
-
N-acetyl-D-glucosamine
pH 8.0, 37°C, recombinant mutant N213A at 50 mM MgCl2
Bifidobacterium longum
16.3
-
ATP
pH 8.0, 37°C, recombinant wild-type enzyme, at 5.0 mM MgCl2
Bifidobacterium longum
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Bifidobacterium longum
General Information
General Information
Commentary
Organism
additional information
nucleotide binding site structures of NahK-ATP and Nahk-ADP, overview
Bifidobacterium longum
General Information (protein specific)
General Information
Commentary
Organism
additional information
nucleotide binding site structures of NahK-ATP and Nahk-ADP, overview
Bifidobacterium longum
Other publictions for EC 2.7.1.162
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)
739523
Li
Efficient chemoenzymatic synth ...
Escherichia coli
Prep. Biochem. Biotechnol.
47
852-859
2017
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
737770
Sato
Open-close structural change u ...
Bifidobacterium longum, Bifidobacterium longum JCM 1217
Biochim. Biophys. Acta
1854
333-340
2015
-
-
1
1
4
-
-
8
-
1
-
2
-
5
-
-
1
1
-
-
-
-
4
2
1
-
-
8
1
-
-
-
-
-
-
-
-
1
-
1
4
-
-
-
-
8
-
1
-
2
-
-
-
1
-
-
-
-
4
2
1
-
-
8
1
-
-
-
-
1
1
-
-
-
737343
Wang
Insights into the binding spec ...
Bifidobacterium longum, Bifidobacterium longum JCM 1217, Bifidobacterium longum subsp. infantis, Bifidobacterium longum subsp. infantis ATCC 15697
Acta Crystallogr. Sect. D
70
1401-1410
2014
-
-
2
2
17
-
-
1
-
2
2
3
-
12
-
-
2
2
-
-
-
-
24
3
2
-
-
-
2
-
-
2
-
-
-
-
-
2
2
2
17
-
-
-
-
1
-
2
2
3
-
-
-
2
-
-
-
-
24
3
2
-
-
-
2
-
-
-
-
4
4
-
-
-
726089
Li
Efficient enzymatic synthesis ...
Bifidobacterium longum subsp. infantis
Org. Lett.
15
5528-5530
2013
-
1
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
723238
Li
Substrate promiscuity of N-ace ...
Bifidobacterium longum, Bifidobacterium longum ATCC 55813, Bifidobacterium longum subsp. infantis, Bifidobacterium longum subsp. infantis ATCC 15697
Molecules
16
6396-6407
2011
-
-
2
-
-
-
-
8
-
4
-
-
-
11
-
-
-
-
-
-
-
-
36
-
-
-
-
8
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
8
-
4
-
-
-
-
-
-
-
-
-
-
36
-
-
-
-
8
2
-
-
-
-
-
-
-
8
8
702588
Cai
Substrate specificity of N-ace ...
Bifidobacterium longum
Bioorg. Med. Chem. Lett.
19
5433-5435
2009
-
-
-
-
-
-
-
-
-
1
-
1
-
1
-
-
-
-
-
-
-
-
11
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
11
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
703179
Cai
A chemoenzymatic route to N-ac ...
Bifidobacterium longum
Chem. Commun. (Camb. )
2009
2944-2946
2009
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
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-
-
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-
-
-
-
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-
-
-
-
1
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
677700
Nishimoto
Identification of N-acetylhexo ...
Bifidobacterium longum, Bifidobacterium longum JCM 1217
Appl. Environ. Microbiol.
73
6444-6449
2007
1
-
1
-
-
1
-
3
-
6
-
6
-
7
-
-
1
1
-
1
13
-
17
-
1
-
1
3
1
-
1
3
-
-
-
1
-
1
3
-
-
1
-
-
-
3
-
6
-
6
-
-
-
1
-
1
13
-
17
-
1
-
1
3
1
-
1
-
-
-
-
-
-
-