title: Switching demethylation activities between AlkB family RNA/DNA demethylases through exchange of active-site residues.
abstract: The AlkB family demethylases AlkB, FTO, and ALKBH5 recognize differentially methylated RNA/DNA substrates, which results in their distinct biological roles. Here we identify key active-site residues that contribute to their substrate specificity. Swapping such active-site residues between the demethylases leads to partially switched demethylation activities. Combined evidence from X-ray structures and enzyme kinetics suggests a role of the active-site residues in substrate recognition. Such a divergent active-site sequence may aid the design of selective inhibitors that can discriminate these homologue RNA/DNA demethylases.
authors.affiliation: State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Synthetic and Functional Biomolecules Center, and Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871 (China).
authors.forename: Chenxu
authors.initials: C
authors.lastname: Zhu
authors.forename: Chengqi
authors.initials: C
authors.lastname: Yi
mesh_terms: D000071499:AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase; D000595:Amino Acid Sequence; D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D045643:DNA Repair Enzymes / Q000235:genetics* / Q000378:metabolism; D049308:Dioxygenases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D012313:RNA / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D012313:RNA; D004247:DNA; D049308:Dioxygenases; C486440:ALKBH2 protein, human; D000071499:AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase; D045643:DNA Repair Enzymes
keywords: RNA recognition; RNA/DNA demethylases; protein structures; selective inhibition; substrate specificity