Cloned (Comment) | Organism |
---|---|
mutants of human Alkbh5 are amplified by PCR and subcloned into a modified pET-28a (Novagen) vector encoding a tobacco etch virus protease recognition site. The final clones are verified by DNA sequencing. All of the recombinant plasmids are transformed into Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
crystallizations are performed at 24 and 4°C using both the hanging drop and sitting drop vapor diffusion methods. Five high resolution crystal structures of the catalytic core of Alkbh5 in complex with different ligands. These findings provide a structural basis for understanding the substrate recognition specificity of Alkbh5 and offer a foundation for selective drug design against AlkB members | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
269A/Q271A | double mutation shows no impact on the repair efficiency of Alkbh5 | Homo sapiens |
F232A/F234A | mutant exhibits 41% activity toward m6A-containing ssDNA | Homo sapiens |
F232D/Q233D/F234E | the mutant enzyme displays a severe loss of activity, demonstrating only 13.5% of wild-type activity | Homo sapiens |
K231A/K235A | double mutation shows no impact on the repair capacity of Alkbh5 | Homo sapiens |
K231E/K235E | double mutation shows no impact on the repair capacity of Alkbh5 | Homo sapiens |
Q146A/K147A/R148A | mutant retains 44% of the wild-type activity | Homo sapiens |
R269E/Q271E | mutant with greatly reduced catalytic activity to 50% | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
citrate | less effective inhibitor | Homo sapiens | |
N-oxalylglycine | - |
Homo sapiens | |
Pyridine 2,4-dicarboxylate | moderate inhibitor | Homo sapiens | |
succinate | - |
Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q6P6C2 | fragment | - |
Purification (Comment) | Organism |
---|---|
- |
Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N6-methyladenine in single-stranded DNA oligonucleotide + 2-oxoglutarate + O2 | - |
Homo sapiens | adenine in single-stranded DNA oligonucleotide + formaldehyde + succinate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ALKBH5 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
21 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.026 | - |
pH 7.2, 21°C | Homo sapiens | N-oxalylglycine | |
0.03 | - |
pH 7.2, 21°C | Homo sapiens | succinate | |
0.347 | - |
pH 7.2, 21°C | Homo sapiens | Pyridine 2,4-dicarboxylate | |
0.628 | - |
pH 7.2, 21°C | Homo sapiens | citrate |
General Information | Comment | Organism |
---|---|---|
metabolism | N6-methylation of adenosine is the most ubiquitous and abundant modification of nucleoside in eukaryotic mRNA and long non-coding RNA. This modification plays an essential role in the regulation of mRNA translation and RNA metabolism | Homo sapiens |