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Literature summary for 2.1.1.72 extracted from

  • Coffin, S.R.; Reich, N.O.
    Escherichia coli DNA adenine methyltransferase: the structural basis of processive catalysis and indirect read-out (2009), J. Biol. Chem., 284, 18390-18400.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli XL2 Blue cells Escherichia coli

Protein Variants

Protein Variants Comment Organism
K139A the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli
N126A the mutant displays decreased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli
N132A the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli
R116A the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli
R137A the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli
R95A the mutant displays increased kcat value compared to the wild type enzyme using the preferred DNA sequence 5'-GCATACCGGATCAAGTAAATG-3' Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000007
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000008
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.00001
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000023
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000027
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000031
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000035
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000044
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000076
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000081
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000083
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.000086
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.00022
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + DNA adenine Escherichia coli Dam methylates the N-6 position of adenine in the DNA sequence 5'-GATC-3' and is highly processive, catalyzing multiple methyltransfers prior to dissociating from the DNA S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0AEE9
-
-

Purification (Commentary)

Purification (Comment) Organism
phosphocellulose column chromatography and Blue Sepharose column chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + DNA adenine Dam methylates the N-6 position of adenine in the DNA sequence 5'-GATC-3' and is highly processive, catalyzing multiple methyltransfers prior to dissociating from the DNA Escherichia coli S-adenosyl-L-homocysteine + DNA 6-methyladenine
-
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Synonyms

Synonyms Comment Organism
Dam
-
Escherichia coli
DNA adenine methyltransferase
-
Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00017
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.00082
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.0009
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.0015
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.0038
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.0055
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.008
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.0083
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.013
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.015
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.019
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.02
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
0.058
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
48.3
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N126A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
100
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
117
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R95A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
125
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
133
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
160
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
167
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
187
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', wild type enzyme, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
223
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
262
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme R137A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
267
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme N132A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
300
-
DNA adenine using the non-preferred sequence 5'-GCATAAAAGATCGTCTAAATG-3', mutant enzyme K139A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli
367
-
DNA adenine using the preferred sequence 5'-GCATACCGGATCAAGTAAATG-3', mutant enzyme R116A, in 20 mM potassium phosphate, pH 7.5, 200 mM NaCl, 0.2 mM EDTA, 0.2 mg/ml bovine serum albumin, 2 mM dithiothreitol, 10% (v/v) glycerol, at 22°C Escherichia coli