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
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
extent of glucosylation varies between 50% and 80% of the amount of hydroxymethylcytosine residues calculated to be present, T4 DNA with no unsubstituted hydroxymethylcytosine groups does not serve as acceptor, T2 DNA: no activity with enzyme from T2-infected cells, small activity with enzyme from T4- and T6-infected cells
UDPglucose + 5-hydroxymethylcytosine containing DNA
UDP + alpha-glucosyl-5-hydroxymethylcytosine containing DNA
Tequatrovirus T4
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extent of glucosylation varies between 50% and 80% of the amount of hydroxymethylcytosine residues calculated to be present, T4 DNA with no unsubstituted hydroxymethylcytosine groups does not serve as acceptor, T2 DNA: no activity with enzyme from T2-infected cells, small activity with enzyme from T4- and T6-infected cells
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E22A
Tequatrovirus T4
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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
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
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
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
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