Literature summary for 2.1.1.72 extracted from

Liebert, K.; Hermann, A.; Schlickenrieder, M.; Jeltsch, A.
Stopped-flow and mutational analysis of base flipping by the Escherichia coli Dam DNA-(adenine-N6)-methyltransferase (2004), J. Mol. Biol., 341, 443-454.

Search for more literature for 2.1.1.72

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D181A	site-directed mutagenesis, inactive mutant, mutation abolishes base flipping, D181 seems to contact and stabilize the flipped base, i.e. the intermediate state of the base flipping process	Escherichia coli
S188A	site-directed mutagenesis, exchange in the loop next to the active site, 7-8fold reduction of kcat, mutant shows 92% of wild-type enzyme activity	Escherichia coli
T190A	site-directed mutagenesis, mutant shows 75% of wild-type enzyme activity	Escherichia coli
Y184A	site-directed mutagenesis, mutant shows 1.7% of wild-type enzyme activity	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	quantitative stopped-flow kinetics using 2-aminopurine as a probe to detect base flipping, wild-type and mutant enzymes	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	-	S-adenosyl-L-homocysteine + DNA 6-methylaminopurine	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes to over 95% purity	Escherichia coli

Reaction

Reaction	Comment	Organism	Reaction ID
S-adenosyl-L-methionine + adenine in DNA = S-adenosyl-L-homocysteine + N6-methyladenine in DNA	D181 and Y184 are essential for activity	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	substrate specificity with different oligonucleotides, substrate binding specificity, overview	Escherichia coli	?	-	?
S-adenosyl-L-methionine + DNA adenine	-	Escherichia coli	S-adenosyl-L-homocysteine + DNA 6-methylaminopurine	-	?
S-adenosyl-L-methionine + DNA adenine	required substrate recognition target sequence is GATC, occuring base flipping in absence of S-adenosyl-L-methionine is a biphasic process and very fast, but binding of the flipped base into the active site pocket requiring S-adenosyl-L-methionine is slow, active site contains the conserved DPPY motif, whose tyrosine184 residue stacks to the flipped target base	Escherichia coli	S-adenosyl-L-homocysteine + DNA 6-methylaminopurine	-	?

Subunits

Subunits	Comment	Organism
More	secondary structure analysis	Escherichia coli

Synonyms

Synonyms	Comment	Organism
Dam DNA-(adenine-N6)-methyltransferase	-	Escherichia coli
Dam DNA-(adenine-N6)-MTase	-	Escherichia coli
EcoDam	-	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	-	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Escherichia coli

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Release 2023.1 (January 2023)