Key Laboratory of Molecular Nanostructure and Nanotechnology, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry , Chinese Academy of Sciences , Beijing 100190 , China.
University of Chinese Academy of Sciences , Beijing 100049 , China.
Anal Chem. 2018 Apr 3;90(7):4596-4602. doi: 10.1021/acs.analchem.7b05070. Epub 2018 Mar 14.
Dimerization of core protein is a crucial step in the formation of the hepatitis C virus (HCV) nucleocapsid, and inhibition of dimer formation is regarded as an attractive approach to design anti-HCV drugs. In this work, we developed the atomic force microscopy based single molecular force spectroscopy (AFM-SMFS) method for the characterization of core protein dimerization with the advantages of small amount of sample consumption and no need of labeling. Interaction force of the core protein with its antibody or aptamer was analyzed to investigate its stoichiometry and binding property. The two specific binding forces were detected due to the probing of dimeric and monomeric core protein, respectively. Moreover, the binding property of protein dimer was different from the monomer. Our work offers a new approach to study the dimerization of core protein, as well as other proteins, and to screen the HCV candidate inhibitors.
核心蛋白二聚化是丙型肝炎病毒（HCV）核衣壳形成的关键步骤，抑制二聚体形成被认为是设计抗 HCV 药物的一种有吸引力的方法。在这项工作中，我们开发了基于原子力显微镜的单分子力谱（AFM-SMFS）方法来表征核心蛋白二聚化，该方法具有样品消耗少和无需标记的优点。分析核心蛋白与其抗体或适体的相互作用力，以研究其化学计量和结合特性。由于分别探测二聚体和单体核心蛋白，因此检测到两个特定的结合力。此外，蛋白质二聚体的结合特性与单体不同。我们的工作为研究核心蛋白以及其他蛋白质的二聚化以及筛选 HCV 候选抑制剂提供了一种新方法。
