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Literature summary extracted from

  • Maynard-Smith, M.; McKelvie, J.; Wood, R.; Harmer, J.; Ranasinghe, R.; Williams, C.; Coomber, D.; Stares, A.; Roach, P.
    Direct and continuous fluorescence-based measurements of Pyrococcus horikoshii DNA N-6 adenine methyltransferase activity (2011), Anal. Biochem., 418, 204-212 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.72
-
 Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.72 M98A to avoid expression of this truncated protein, a mutant form of the Pyrococcus horikoshii dam gene (M98A) is prepared by site-directed mutagenesis Pyrococcus horikoshii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.72 0.00000191
-
 DNA adenine pH 7.8, 33°C Pyrococcus horikoshii
2.1.1.72 0.000396
-
 S-adenosyl-L-methionine pH 7.8, 33°C Pyrococcus horikoshii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.72 40200
-
 SDS-PAGE Pyrococcus horikoshii

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.72 Pyrococcus horikoshii
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.72
-
 Pyrococcus horikoshii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.72 additional information
-
 the enzyme catalyzes the methylation of adenine at the N-6 position within its DNA recognition sequence GATC. Enzymatic methylation of the hemimethylated GATC site results in destabilisation of the duplex, increasing the proportion of dissociated DNA, and producing an observable increase in fluorescence. By using a suitable oligonucleotide substrate, labeled on adjacent strands with a fluorophore (fluorescein) and quencher (dabcyl), the destabilization of duplex DNA due to adenine methylation can be monitored by fluorescence measurements. The separation of fluorophore and quencher during strand dissociation causes an observable increase in fluorescence, providing a reproducible, direct, and real-time activity assay Pyrococcus horikoshii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.72 S-adenosyl-L-methionine + DNA adenine the enzyme catalyzes the methylation of adenine at the N-6 position within its DNA recognition sequence GATC. Enzymatic methylation of the hemimethylated GATC site results in destabilisation of the duplex, increasing the proportion of dissociated DNA, and producing an observable increase in fluorescence. By using a suitable oligonucleotide substrate, labeled on adjacent strands with a fluorophore (fluorescein) and quencher (dabcyl), the destabilization of duplex DNA due to adenine methylation can be monitored by fluorescence measurements. The separation of fluorophore and quencher during strand dissociation causes an observable increase in fluorescence, providing a reproducible, direct, and real-time activity assay Pyrococcus horikoshii S-adenosyl-L-homocysteine + DNA 6-methylaminopurine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.72 ? x * 40200, SDS-PAGE Pyrococcus horikoshii

Synonyms

EC Number Synonyms Comment Organism
2.1.1.72 DNA N-6 adenine methyltransferase
-
 Pyrococcus horikoshii
2.1.1.72 Pho(M98A)Dam
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.72 33
-
 assay at Pyrococcus horikoshii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.72 0.004
-
 S-adenosyl-L-methionine pH 7.8, 33°C Pyrococcus horikoshii
2.1.1.72 0.0042
-
 DNA adenine pH 7.8, 33°C Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.72 7.8
-
 assay at Pyrococcus horikoshii

General Information

EC Number General Information Comment Organism
2.1.1.72 metabolism the enzyme plays a critical role in the regulation of cellular processes within many bacterial species, including mismatch repair and the timing of DNA replication Pyrococcus horikoshii