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Literature summary for 2.1.1.72 extracted from

  • Bheemanaik, S.; Chandrashekaran, S.; Nagaraja, V.; Rao, D.N.
    Kinetic and catalytic properties of dimeric KpnI DNA methyltransferase (2003), J. Biol. Chem., 278, 7863-7874.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli strain DH10B Klebsiella pneumoniae

Inhibitors

Inhibitors Comment Organism Structure
methylated duplex DNA partial inhibition at high concentrations, mixed inhibition against un- or hemimethylated DNA, noncompetitive against S-adenosyl-L-methionine Klebsiella pneumoniae
S-adenosyl-L-homocysteine competitive inhibition against S-adenosyl-L-methionine, noncompetitive against DNA Klebsiella pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics, ordered bi bi kinetic mechanism, cooperative binding of 2 molecules of enzyme per DNA is required for activity Klebsiella pneumoniae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
2 * 45000, recombinant enzyme, SDS-PAGE Klebsiella pneumoniae
90000
-
recombinant enzyme, gel filtration Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + DNA adenine Klebsiella pneumoniae enzyme is a member of a restriction-modification system, R-M system, plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
?

Organism

Organism UniProt Comment Textmining
Klebsiella pneumoniae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain DH10B to homogeneity by ultracentrifugation, treatment by 1% v/v polyethyleneimine with 250 mM KCl, ammonium sulfate fractionation, several steps of dialysis, phosphocellulose resin chromatography, and affinity chromatography Klebsiella pneumoniae

Reaction

Reaction Comment Organism Reaction ID
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA ordered bi bi mechanism with S-adenosyl-L-homocysteine leaving at last, cooperative binding of 2 molecules of enzyme per DNA is required for activity Klebsiella pneumoniae

Storage Stability

Storage Stability Organism
-20°C, purified recombinant enzyme, 10 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 100 mM KCl, 7 mM 2-mercaptoethanol, and 10% glycerol, stable for at least 3 months Klebsiella pneumoniae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + adenine in DNA
-
Klebsiella pneumoniae S-adenosyl-L-homocysteine + N6-methyladenine in DNA
-
r
S-adenosyl-L-methionine + DNA adenine enzyme is a member of a restriction-modification system, R-M system, plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA Klebsiella pneumoniae S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
?
S-adenosyl-L-methionine + DNA adenine enzyme recognizes the sequence 5'-GGTACC-3', methylation at position N6, enzyme utilizes plasmid DNA, and hemi- or unmethylated duplex DNA, preference for hemimethylated DNA Klebsiella pneumoniae S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
?

Subunits

Subunits Comment Organism
dimer 2 * 45000, recombinant enzyme, SDS-PAGE Klebsiella pneumoniae

Synonyms

Synonyms Comment Organism
KpnI DNA methyltransferase
-
Klebsiella pneumoniae
KpnI DNA-(N6-adenine)-methyltransferase
-
Klebsiella pneumoniae
KpnI MTase
-
Klebsiella pneumoniae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Klebsiella pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00033
-
adenine in DNA substrate plasmid DNA, recombinant enzyme, pH 8.0, 37°C Klebsiella pneumoniae
0.0011
-
adenine in DNA substrate unmethylated duplex DNA, recombinant enzyme, pH 8.0, 37°C Klebsiella pneumoniae
0.0056
-
adenine in DNA substrate hemimethylated duplex DNA, recombinant enzyme, pH 8.0, 37°C Klebsiella pneumoniae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Klebsiella pneumoniae

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
additional information inhibition kinetics Klebsiella pneumoniae
0.00062
-
S-adenosyl-L-homocysteine versus S-adenosyl-L-methionine, pH 8.0, 37°C Klebsiella pneumoniae