Literature summary for 2.1.1.72 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 .

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Cloned(Commentary)

Cloned (Comment)	Organism
-	Pyrococcus horikoshii

Protein Variants

Protein Variants	Comment	Organism
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]

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

Molecular Weight [Da]

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

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus horikoshii	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Pyrococcus horikoshii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
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)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
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

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

Synonyms

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

General Information

General Information	Comment	Organism
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

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Release 2023.1 (January 2023)