Literature summary for 2.4.1.26 extracted from

Sommer, N.; Depping, R.; Piotrowski, M.; Ruger, W.
Bacteriophage T4 alpha-glucosyltransferase: a novel interaction with gp45 and aspects of the catalytic mechanism (2004), Biochem. Biophys. Res. Commun., 323, 809-815.

Search for more literature for 2.4.1.26

Cloned(Commentary)

Cloned (Comment)	Organism
gene agt, overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain XL1	Tequatrovirus T4

Protein Variants

Protein Variants	Comment	Organism
E22A	site-directed mutagenesis, 50% activity compared to the wild-type enzyme, in absence of acceptor 70% reduced UDG-glucose turnover compared to the wild-type enzyme	Tequatrovirus T4
E311A	site-directed mutagenesis, 1-2% activity compared to the wild-type enzyme, in absence of acceptor no remaining UDG-glucose turnover	Tequatrovirus T4
M231A	site-directed mutagenesis, 1-2% activity compared to the wild-type enzyme, in absence of acceptor mutant shows an unaltered UDG-glucose turnover compared to the wild-type enzyme	Tequatrovirus T4
R236A	site-directed mutagenesis, 27% activity compared to the wild-type enzyme, in absence of acceptor mutant shows an unaltered UDG-glucose turnover compared to the wild-type enzyme	Tequatrovirus T4
R256A	site-directed mutagenesis, activity is similar to the wild-type enzyme, in absence of acceptor mutant shows an unaltered UDG-glucose turnover compared to the wild-type enzyme	Tequatrovirus T4
S106A	site-directed mutagenesis, 111% activity compared to the wild-type enzyme, in absence of acceptor mutant shows an unaltered UDG-glucose turnover compared to the wild-type enzyme	Tequatrovirus T4

Organism

Organism	UniProt	Comment	Textmining
Tequatrovirus T4	-	gene agt	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain XL1 by metal affinity chromatography	Tequatrovirus T4

Reaction

Reaction	Comment	Organism	Reaction ID
UDPglucose + 5-hydroxymethylcytosine containing DNA = UDP + alpha-glucosyl-5-hydroxymethylcytosine containing DNA	catalytic mechanism, DNA binding and base flipping mechanism	Tequatrovirus T4	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	hydroxymethylated DNA binding activities of wild-type and mutant enzymes, the enzyme undergoes a conformational change from a substrate-free open form to a closed substrate-binding form stabilized by several salt bridges and probably involving Ser106 and Glu22	Tequatrovirus T4	?	-	?
UDP-glucose + hydroxymethylated DNA	Glu22 seems not to be responsible for activation of the nucleophilic attack, Ser106 and Glu311 probably form a salt bridge in the closed enzyme formation, Glu311 is probably involved in interaction with UDP-glucose, Arg236, Arg256 and Met231 are probably involved in interaction with the DNA	Tequatrovirus T4	UDP + glycosylated hydroxymethylated DNA	-	?

Subunits

Subunits	Comment	Organism
More	secondary structure alignment	Tequatrovirus T4

Synonyms

Synonyms	Comment	Organism
AGT	-	Tequatrovirus T4

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Tequatrovirus T4

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Tequatrovirus T4

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Release 2023.1 (January 2023)