Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT, 06030, USA.
Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, 02115, USA.
Biomol NMR Assign. 2024 Dec;18(2):293-297. doi: 10.1007/s12104-024-10202-5. Epub 2024 Sep 13.
The 81 kDa E. coli β clamp is a ring-shaped head-to-tail homodimer that encircles DNA and plays a central role in bacterial DNA replication by serving as a processivity factor for DNA polymerases and a binding platform for other DNA replication and repair proteins. Here we report the backbone H, N, and C NMR resonance assignments of the stabilized T45R/S107R β clamp variant obtained using standard TROSY-based triple-resonance experiments (BMRB 52548). The backbone assignments were aided by C and N edited NOESY experiments, allowing us to utilize our previously reported assignments of the β clamp ILV side-chain methyl groups (BMRB 51430, 51431). The backbone assignments of the T45R/S107R β clamp variant were transferred to the wild-type β clamp using a minimal set of TROSY-based N edited NOESY, NHCO and NHCA experiments (BMRB 52549). The reported backbone and previous ILV side-chain resonance assignments will enable NMR studies of the β clamp interactions and dynamics using amide and methyl groups as probes.
81 kDa 的大肠杆菌β 夹是一种环形的头到尾同二聚体，它环绕 DNA，并通过作为 DNA 聚合酶的持续合成因子和其他 DNA 复制和修复蛋白的结合平台，在细菌 DNA 复制中发挥核心作用。在这里，我们报告了使用标准 TROSY 三重共振实验（BMRB 52548）获得的稳定的 T45R/S107R β 夹变体的骨架 H、N 和 C NMR 共振分配。骨架分配得到了 C 和 N 编辑的 NOESY 实验的辅助，允许我们利用我们之前报告的 β 夹 ILV 侧链甲基基团的分配（BMRB 51430、51431）。T45R/S107R β 夹变体的骨架分配通过使用一组最小的基于 TROSY 的 N 编辑 NOESY、NHCO 和 NHCA 实验（BMRB 52549）转移到野生型 β 夹上。报告的骨架和以前的 ILV 侧链共振分配将使酰胺和甲基基团作为探针的 β 夹相互作用和动力学的 NMR 研究成为可能。
