BRENDA - Enzyme Database show

Structural basis of substrate recognition in human nicotinamide N-methyltransferase

Peng, Y.; Sartini, D.; Pozzi, V.; Wilk, D.; Emanuelli, M.; Yee, V.C.; Biochemistry 50, 7800-7808 (2011)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.1.1
expressed in Escherichia coli BL21(DE3)
Homo sapiens
Crystallization (Commentary)
EC Number
Crystallization
Organism
2.1.1.1
His-tagged K100A/E101A/E103A mutant, sitting drop vapor diffusion method, PEG3350
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.1
D197A
active site residue
Homo sapiens
2.1.1.1
K100A/E101A/E103A
His-tagged version, crystallization facilitated by reducing the entropy of three surface exposed residues
Homo sapiens
2.1.1.1
S201A
active site residue
Homo sapiens
2.1.1.1
S213A
active site residue
Homo sapiens
2.1.1.1
Y20A
active site residue
Homo sapiens
2.1.1.1
Y20F
active site residue
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.1
0.002
-
S-adenosyl-L-methionine
0.0018-0.0022 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.005
-
S-adenosyl-L-methionine
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.028
-
S-adenosyl-L-methionine
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.105
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.24
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.4
-
nicotinamide
0.43-0.38 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.63
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
2.73
-
S-adenosyl-L-methionine
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
3.5
-
S-adenosyl-L-methionine
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
11.8
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.1
Homo sapiens
P40261
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.1.1
His-tagged K100A/E101A/E103A mutant, immobilized metal ion affinity chromatography (Ni2+), gel filtration
Homo sapiens
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.1
0.0011
-
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.0042
-
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.077
-
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.14
-
S201A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.15
-
wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.18
-
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.21
-
S213A mutant protein, pH 8.6, 37°C
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.1
nicotinamide + S-adenosyl-L-methionine
-
718914
Homo sapiens
1-methylnicotinamide + S-adenosyl-L-homocysteine
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.1
0.031
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.036
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.041
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.077
-
nicotinamide
0.43-0.38 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.083
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.1
expressed in Escherichia coli BL21(DE3)
Homo sapiens
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.1.1.1
His-tagged K100A/E101A/E103A mutant, sitting drop vapor diffusion method, PEG3350
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.1.1.1
D197A
active site residue
Homo sapiens
2.1.1.1
K100A/E101A/E103A
His-tagged version, crystallization facilitated by reducing the entropy of three surface exposed residues
Homo sapiens
2.1.1.1
S201A
active site residue
Homo sapiens
2.1.1.1
S213A
active site residue
Homo sapiens
2.1.1.1
Y20A
active site residue
Homo sapiens
2.1.1.1
Y20F
active site residue
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.1
0.002
-
S-adenosyl-L-methionine
0.0018-0.0022 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.005
-
S-adenosyl-L-methionine
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.028
-
S-adenosyl-L-methionine
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.105
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.24
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.4
-
nicotinamide
0.43-0.38 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.63
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
2.73
-
S-adenosyl-L-methionine
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
3.5
-
S-adenosyl-L-methionine
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
11.8
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.1
His-tagged K100A/E101A/E103A mutant, immobilized metal ion affinity chromatography (Ni2+), gel filtration
Homo sapiens
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.1
0.0011
-
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.0042
-
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.077
-
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.14
-
S201A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.15
-
wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.18
-
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.21
-
S213A mutant protein, pH 8.6, 37°C
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.1
nicotinamide + S-adenosyl-L-methionine
-
718914
Homo sapiens
1-methylnicotinamide + S-adenosyl-L-homocysteine
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.1
0.031
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.036
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.041
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.077
-
nicotinamide
0.43-0.38 mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.083
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.1.1.1
0.003
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.005
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.009
-
S-adenosyl-L-methionine
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.013
-
S-adenosyl-L-methionine
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.17
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.19
-
nicotinamide
0.18-0.2 1/sec*mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.8
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
1.5
-
S-adenosyl-L-methionine
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
17
-
S-adenosyl-L-methionine
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
39
-
S-adenosyl-L-methionine
35-43 1/sec*mM, wild type protein, pH 8.6, 37°C
Homo sapiens
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
2.1.1.1
0.003
-
nicotinamide
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.005
-
nicotinamide
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.009
-
S-adenosyl-L-methionine
Y20A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.013
-
S-adenosyl-L-methionine
D197A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.17
-
nicotinamide
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.19
-
nicotinamide
0.18-0.2 1/sec*mM, wild type protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
0.8
-
nicotinamide
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
1.5
-
S-adenosyl-L-methionine
Y20F mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
17
-
S-adenosyl-L-methionine
K100A/E101A/E103A mutant protein, pH 8.6, 37°C
Homo sapiens
2.1.1.1
39
-
S-adenosyl-L-methionine
35-43 1/sec*mM, wild type protein, pH 8.6, 37°C
Homo sapiens