Li Yan, Phoo Wint Wint, Loh Ying Ru, Zhang Zhenzhen, Ng Elizabeth Yihui, Wang Weiling, Keller Thomas H, Luo Dahai, Kang CongBao
Experimental Therapeutics Centre Agency for Science Technology and Research (A*STAR), Singapore, Singapore.
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore.
FEBS Lett. 2017 Aug;591(15):2338-2347. doi: 10.1002/1873-3468.12741. Epub 2017 Jul 20.
The Zika virus (ZIKV) NS2B-NS3 protease is an important drug target. The conventional flaviviral protease constructs used for structural studies contain the NS2B cofactor region linked to the NS3 protease domain via a glycine-rich flexible linker. Here, we examined the structural dynamics of this conventional Zika protease (gZiPro) using NMR spectroscopy. Although the glycine-rich linker in gZiPro does not alter the overall folding of the protease in solution, gZiPro is not homogenous in ion exchange chromatography. Compared to the unlinked protease construct, the artificial linker affects the chemical environment of many residues including H51 in the catalytic triad. Our study provides a direct comparison of ZIKV protease constructs with and without an artificial linker.
寨卡病毒（ZIKV）NS2B-NS3蛋白酶是一个重要的药物靶点。用于结构研究的传统黄病毒蛋白酶构建体包含通过富含甘氨酸的柔性接头与NS3蛋白酶结构域相连的NS2B辅助因子区域。在此，我们使用核磁共振光谱研究了这种传统寨卡蛋白酶（gZiPro）的结构动力学。尽管gZiPro中富含甘氨酸的接头不会改变蛋白酶在溶液中的整体折叠，但gZiPro在离子交换色谱中并非均一。与无接头的蛋白酶构建体相比，人工接头影响包括催化三联体中H51在内的许多残基的化学环境。我们的研究对有无人工接头的寨卡病毒蛋白酶构建体进行了直接比较。
