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1.14.11.53: mRNA N6-methyladenine demethylase

This is an abbreviated version!
For detailed information about mRNA N6-methyladenine demethylase, go to the full flat file.

Word Map on EC 1.14.11.53

Reaction

N6-methyladenine in mRNA
+
2-oxoglutarate
 +
O2
=
adenine in mRNA
 +
formaldehyde
 +
succinate
 +
CO2

Synonyms

AlkB homolog 5, ALKBH10B, ALKBH5, ALKBH5 demethylase, ALKBH9B, alkylation repair homolog protein 5, fat mass and obesity-associated enzyme, fat mass and obesity-associated protein, FTO, m6A mRNA demethylase, m6A RNA demethylase, m6A-RNA demethylase, N6-methyladenosine demethylase, RNA N6-methyladenine demethylase

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.11 With 2-oxoglutarate as one donor, and incorporation of one atom of oxygen into each donor
                1.14.11.53 mRNA N6-methyladenine demethylase

Crystallization

Crystallization on EC 1.14.11.53 - mRNA N6-methyladenine demethylase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with manganese(II) and 2-oxoglutarate, sitting drop vapor diffusion method, using 0.2 M sodium iodide, pH 7.0, 20% (w/v) polyethylene glycol 3350
ALKBH566–292 is crystallized in sitting drops at 20°C by the vapour diffusion method in the presence of Mn2+ and (1-chloro-4-hydroxyisoquinoline-3-carbonyl)glycine. Crystallization drops contain 0.2 ml of a protein solution containing a final concentration of 10 mg/ml hexahistidine-tagged ALKBH566–292, 0.5 mM MnCl2 and 2 mM IOX3 mixed with 0.1 ml of well solution containing 125 mM potassium nitrate and 15% (w/v) polyethylene glycol 3350. Crystals (size 100 x 50 x 50 mM) appeared after 3 months. Crystals are harvested using nylon loops and cryoprotected using well solution diluted with 25% (v/v) glycerol and flashcooled in liquid nitrogen. Crystal structure of human ALKBH5 (residues 66-292) to 2.0 A resolution. ALKBH566–292 has a double-stranded beta-helix core fold. The active site metal is octahedrally coordinated by an HXD...H motif (comprising residues His204, Asp206 and His266) and three water molecules. ALKBH5 shares a nucleotide recognition lid and conserved active site residues with other ferrous iron-dependent nucleic acid oxygenase
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
hanging drop vapor diffusion method at 18 °C. The ALKBH5–2-oxoglutarate-Mn2+ is crystallized in a buffer containing 0.2 M ammonium dihydrogen phosphate, 20% PEG 3350. The ALKBH5 is crystallized with citrate in a buffer containing ammonium citrate, 20% PEG 3350. Before flashfreezing crystals in liquid nitrogen, crystals are soaked in a cryoprotectant consisting mother liquor plus 12% glycerol
hanging drop vapor diffusion method, using 0.2 M ammonium citrate dibasic pH 5.1, 20% (w/v) PEG 3350
-
hanging-drop vapour-diffusion method, crystal structure of FTO in complex with the mononucleotide 3-methylthymine