nmr analysis of the dynamic exchange of the ns2b cofactor between open and closed conformations of the west nile virus ns2b-ns3 protease核磁共振分析的动态交换ns2b开启和关闭之间的代数余子式构象的西尼罗河病毒ns2b-ns3蛋白酶.pdfVIP

NMR Analysis of the Dynamic Exchange of the NS2B Cofactor between Open and Closed Conformations of the West Nile Virus NS2B-NS3 Protease 1. 1. 1 2¤ 2 1 Xun-Cheng Su , Kiyoshi Ozawa , Ruhu Qi , Subhash G. Vasudevan , Siew P. Lim , Gottfried Otting * 1 Australian National University, Research School of Chemistry, Canberra, Australia, 2 Novartis Institute for Tropical Diseases, Chromos, Singapore Abstract Background: The two-component NS2B-NS3 proteases of West Nile and dengue viruses are essential for viral replication and established targets for drug development. In all crystal structures of the proteases to date, the NS2B cofactor is located far from the substrate binding site (open conformation) in the absence of inhibitor and lining the substrate binding site (closed conformation) in the presence of an inhibitor. Methods: In this work, nuclear magnetic resonance (NMR) spectroscopy of isotope and spin-labeled samples of the West Nile virus protease was used to investigate the occurrence of equilibria between open and closed conformations in solution. Findings: In solution, the closed form of the West Nile virus protease is the predominant conformation irrespective of the presence or absence of inhibitors. Nonetheless, dissociation of the C-terminal part of the NS2B cofactor from the NS3 protease (open conformation) occurs in both the presence and the absence of inhibitors. Low-molecular-weight inhibitors can shift the conformational exchange equilibria so that over 90% of the West Nile virus protease molecules assume the closed conformation. The West Nile virus protease differs from the dengue virus protease, where the open conformation is the predominant form in the absence