1H, 15N, 13C backbone resonance assignment of Escherichia coli AlkB in complex with Zn(II) and α-ketoglutarate
TL;DR
The backbone 1H, 15N, 13C chemical shift assignment of E. coli AlkB in complex with Zn2+ and α-ketoglutarate is reported, allowing investigations of the protein structural dynamics anticipated to provide crucial insight regarding fundamental aspects in the recognition and enzyme regulation processes.
Abstract
Protein and nucleic acid alkylation are important genetic and epigenetic modifications. The dynamic balance between alkylation and dealkylation is regulated by distinct sets of enzymes and is essential for maintaining genomic stability. Escherichia coli AlkB is a member of the Alkylation B (AlkB) family of dioxygenases that dealkylates a wide range of nucleic acid substrates in E. coli, thereby playing a crucial role in cellular repair processes and epigenetic regulation. Here, we report the backbone 1H, 15N, 13C chemical shift assignment of E. coli AlkB in complex with Zn2+ and α-ketoglutarate. Experiments were acquired at 20 °C by heteronuclear multidimensional NMR spectroscopy. Collectively, 91% of all 13C, 15N and 1H backbone resonances of Alkb were assigned, with 182 out of a possible 200 residues assigned in the 1H–15N TROSY spectrum. Using the program TALOS + , a secondary structure prediction was generated from the assigned backbone resonances that is consistent with the previously reported X-ray structure of the enzyme. The reported assignment will permit investigations of the protein structural dynamics anticipated to provide crucial insight regarding fundamental aspects in the recognition and enzyme regulation processes.