Faal Baboucarr, Purslow Jeffrey A, Venditti Vincenzo
Department of Chemistry, Iowa State University, Hach Hall, 2438 Pammel Drive, Ames, IA, 50011, USA.
Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA, 50011, USA.
Biomol NMR Assign. 2025 Jun;19(1):65-69. doi: 10.1007/s12104-025-10219-4. Epub 2025 Jan 29.
The Alkbh7 protein, a member of the Alkylation B (AlkB) family of dioxygenases, plays a crucial role in epigenetic regulation of cellular metabolism. This paper focuses on the NMR backbone resonance assignment of Alkbh7, a fundamental step in understanding its three-dimensional structure and dynamic behavior at the atomic level. Herein, we report the backbone H, N, C chemical shift assignment of the full-length human Alkbh7. Experiments were acquired at 25 °C by heteronuclear multidimensional NMR spectroscopy. Collectively, 70% of the backbone NH resonances were assigned, with 144 out of a possible 205 residues assigned in the H-N TROSY spectrum. Interestingly, peaks from the active site and the C-terminal end of Alkbh7 are not NMR visible, suggesting that these regions are dynamic on the intermediate exchange regime. Using the program TALOS+, a secondary structure prediction was generated from the assigned backbone resonance 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.
Alkbh7蛋白是双加氧酶烷基化B（AlkB）家族的成员，在细胞代谢的表观遗传调控中起着关键作用。本文重点关注Alkbh7的核磁共振主链共振归属，这是在原子水平上理解其三维结构和动态行为的基本步骤。在此，我们报告了全长人Alkbh7的主链H、N、C化学位移归属。实验在25°C下通过异核多维核磁共振光谱法进行。总共归属了70%的主链NH共振，在H-N TROSY谱中，205个可能的残基中有144个被归属。有趣的是，Alkbh7活性位点和C末端的峰在核磁共振中不可见，这表明这些区域在中间交换机制上是动态的。使用TALOS+程序，根据已归属的主链共振生成了二级结构预测，该预测与先前报道的该酶的X射线结构一致。所报告的归属将允许对蛋白质结构动力学进行研究，预计这将为识别和酶调节过程的基本方面提供关键见解。
