Department of Molecular Biology and Biophysics, University of Connecticut Health Center, 06030, Farmington, CT, USA.
Department of Chemistry and Chemical Biology, Northeastern University, 02115, Boston, MA, USA.
Biomol NMR Assign. 2022 Oct;16(2):317-323. doi: 10.1007/s12104-022-10097-0. Epub 2022 Jun 10.
The ring-shaped E. coli β-clamp protein is an 81 kDa head-to-tail homodimer, which serves as a processivity factor anchoring the replicative polymerase to DNA, thereby increasing replication processivity and speed. In addition, it facilitates numerous protein transactions that take place on DNA during replication, repair, and damage response. We used a structure-based approach to obtain nearly complete Ile, Leu and Val side-chain methyl NMR resonance assignments of the wild-type β-clamp and its stabilized T45R/S107R variant based on site directed mutagenesis and the analysis of methyl-methyl NOESY data. The obtained assignments will facilitate future studies of the β-clamp interactions and dynamics.
环形大肠杆菌β-夹蛋白是一个 81 kDa 的头对头同二聚体，作为一个持续因子将复制聚合酶锚定在 DNA 上，从而增加复制的持续性和速度。此外，它还促进了许多在复制、修复和损伤反应过程中发生在 DNA 上的蛋白质转化。我们使用基于结构的方法，通过定点突变和甲基-甲基 NOESY 数据的分析，获得了野生型β-夹及其稳定化 T45R/S107R 变体的几乎完整的异亮氨酸、亮氨酸和缬氨酸侧链甲基 NMR 共振分配。获得的分配将有助于未来对β-夹相互作用和动力学的研究。
