Cloned (Comment) | Organism |
---|---|
gene agt, overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain XL1 | Tequatrovirus T4 |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Tequatrovirus T4 | - |
gene agt | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain XL1 by metal affinity chromatography | Tequatrovirus T4 |
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 |
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 | Comment | Organism |
---|---|---|
More | secondary structure alignment | Tequatrovirus T4 |
Synonyms | Comment | Organism |
---|---|---|
AGT | - |
Tequatrovirus T4 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Tequatrovirus T4 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Tequatrovirus T4 |