College of Chemistry and Chemical Engineering, Fujian Provincial Key Laboratory of Chemical Biology, Xiamen University, Xiamen, China.
State Key Laboratory for Cellular Stress Biology, Department of Biomedical Sciences, School of Life Sciences, Xiamen University, Xiang'an, Xiamen, China.
Protein Sci. 2019 Sep;28(9):1720-1726. doi: 10.1002/pro.3687.
Lon protease is evolutionarily conserved in prokaryotes and eukaryotic organelles. The primary function of Lon is to selectively degrade abnormal and certain regulatory proteins to maintain the homeostasis in vivo. Lon mainly consists of three functional domains and the N-terminal domain is required for the substrate selection and recognition. However, the precise contribution of the N-terminal domain remains elusive. Here, we determined the crystal structure of the N-terminal 192-residue construct of Lon protease from Mycobacterium avium complex at 2.4 å resolution,and measured NMR-relaxation parameters of backbones. This structure consists of two subdomains, the β-strand rich N-terminal subdomain and the five-helix bundle of C-terminal subdomain, connected by a flexible linker,and is similar to the overall structure of the N domain of Escherichia coli Lon even though their sequence identity is only 26%. The obtained NMR-relaxation parameters reveal two stabilized loops involved in the structural packing of the compact N domain and a turn structure formation. The performed homology comparison suggests that structural and sequence variations in the N domain may be closely related to the substrate selectivity of Lon variants. Our results provide the structure and dynamics characterization of a new Lon N domain, and will help to define the precise contribution of the Lon N-terminal domain to the substrate recognition.
Lon 蛋白酶在原核生物和真核细胞器中是进化保守的。Lon 的主要功能是选择性降解异常和某些调节蛋白,以维持体内的动态平衡。Lon 主要由三个功能域组成,N 端结构域负责底物的选择和识别。然而,N 端结构域的确切贡献仍不清楚。在这里,我们测定了来自鸟分枝杆菌复合体的 Lon 蛋白酶的 N 端 192 个残基结构的晶体结构,分辨率为 2.4 å,测量了骨架的 NMR 弛豫参数。该结构由富含 β-链的 N 端结构域和 C 端五螺旋束结构域组成,通过一个柔性连接子连接,与大肠杆菌 Lon 的 N 结构域的整体结构相似,尽管它们的序列同一性仅为 26%。获得的 NMR 弛豫参数揭示了两个稳定的环,涉及紧凑的 N 结构域的结构包装和转角结构的形成。进行的同源性比较表明,N 结构域的结构和序列变化可能与 Lon 变体的底物选择性密切相关。我们的研究结果提供了一个新的 Lon N 结构域的结构和动力学特征,将有助于确定 Lon N 端结构域在底物识别中的精确贡献。