Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, Missouri 65212, United States.
Department of Biochemistry, University of Missouri, Columbia, Missouri 65211, United States.
ACS Infect Dis. 2024 Aug 9;10(8):2637-2655. doi: 10.1021/acsinfecdis.3c00708. Epub 2024 Jul 17.
The HIV-1 capsid protein (CA) assumes distinct structural forms during replication, each presenting unique, solvent-accessible surfaces that facilitate multifaceted functions and host factor interactions. However, functional contributions of individual CA structures remain unclear, as evaluation of CA presents several technical challenges. To address this knowledge gap, we identified CA-targeting aptamers with different structural specificities, which emerged through a branched SELEX approach using an aptamer library previously selected to bind the CA hexamer lattice. Subsets were either highly specific for the CA lattice or bound both the CA lattice and CA hexamer. We then evaluated four representatives to reveal aptamer regions required for binding, highlighting interesting structural features and challenges in aptamer structure determination. Further, we demonstrate binding to biologically relevant CA structural forms and aptamer-mediated affinity purification of CA from cell lysates without virus or host modification, supporting the development of structural form-specific aptamers as exciting new tools for the study of CA.
HIV-1 衣壳蛋白 (CA) 在复制过程中呈现出不同的结构形式，每种形式都具有独特的、可溶剂化的表面，从而促进多方面的功能和宿主因子相互作用。然而，由于评估 CA 存在几个技术挑战，因此个别 CA 结构的功能贡献仍不清楚。为了解决这一知识空白，我们使用先前选择用于结合 CA 六聚体晶格的适体文库，通过分支 SELEX 方法鉴定了具有不同结构特异性的 CA 靶向适体。这些适体子集要么高度特异性地结合 CA 晶格，要么同时结合 CA 晶格和 CA 六聚体。然后，我们评估了四个代表物，以揭示结合所需的适体区域，突出了适体结构确定中的有趣结构特征和挑战。此外，我们证明了与生物相关的 CA 结构形式的结合以及在不改变病毒或宿主的情况下从细胞裂解物中通过适体介导的 CA 亲和纯化，支持了结构形式特异性适体的开发，这些适体是研究 CA 的令人兴奋的新工具。
