Real-time observation of native conformations and molecular behaviors of viral fusion proteins using high-speed atomic force microscopy (HS-AFM)

使用高速原子力显微镜 (HS-AFM) 实时观察病毒融合蛋白的天然构象和分子行为

• 批准号: 20K16262
• 负责人: LIM KEE・SIANG
• 金额: $ 2.66万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2020
• 资助国家: 日本
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-20K16262/
• 关键词: SARS-CoV-2 spike protein; High-speed AFM; Neutralizing antibodies; COVID-19; HS-AFM; Viral Fusion Protein; Viral fusion protein; fusogenic transition; exosome; Direct visualization; Virulence factors

The objectives of this project are: 1) to record the native conformation viral fusion proteins under physiological conditions using HS-AFM; 2) to visualize the rapid and transient intermediates during conformational changes triggered by acidic conditions using HS-AFM; 3) to elucidate the conformational dynamic of viral fusion proteins when interacting with exosomes or antibodies using HS-AFM. The PI has implemented HS-AFM to perform nanoscopic elucidation of SARS-CoV-2 spike (S) protein structural dynamics during its interaction with neutralizing antibodies (Objective 3). Certain neutralizing antibodies promote the transition of RBD from its close conformation to open confirmation to enhance RBD-ACE2 interaction. This effect is known as antibody-dependent enhancement effect (ADE). This study reveals that HS-AFM could be worked as an efficient nanoscopic assessment platform for S NAb testing. This study has been published in a well-reputed journal, Nano Letters. Furthermore, this study has also been presented in several domestic conferences. Envelope protein (E protein) of Dengue virus will be investigated in the near future.

本课题的目的是：1)利用HS-AFM记录生理条件下的天然构象病毒融合蛋白；2)利用HS-AFM可视化酸性条件引发构象变化过程中的快速和瞬态中间体；3)利用HS-AFM阐明病毒融合蛋白与外泌体或抗体相互作用时的构象动态。PI采用HS-AFM对SARS-CoV-2与中和抗体相互作用过程中的刺突(S)蛋白结构动力学进行纳米级解析（目的3）。某些中和抗体促进RBD由封闭构象向开放构象转变，从而增强RBD与ace2的相互作用。这种效应被称为抗体依赖性增强效应（ADE）。本研究表明，HS-AFM可以作为一种高效的S- NAb检测的纳米级评价平台。这项研究发表在著名杂志《纳米快报》上。此外，该研究还在国内多个会议上发表。登革热病毒的包膜蛋白（E蛋白）将在不久的将来得到进一步的研究。

项目成果

Nanoscopic Assessment of Anti-SARS-CoV-2 Spike Neutralizing Antibody Using High-Speed AFM.

• DOI: 10.1021/acs.nanolett.2c04270
• 发表时间: 2023-01-25
• 期刊: NANO LETTERS
• 影响因子: 10.8
• 作者: Lim, Keesiang;Nishide, Goro;Sajidah, Elma Sakinatus;Yamano, Tomoyoshi;Qiu, Yujia;Yoshida, Takeshi;Kobayashi, Akiko;Hazawa, Masaharu;Ando, Toshio;Hanayama, Rikinari;Wong, Richard W.
• 通讯作者: Wong, Richard W.

Structural dynamics of SARS-CoV-2 spike (S) protein and its interaction with ACE2 receptor and small extracellular vesicle

SARS-CoV-2刺突（S）蛋白的结构动力学及其与ACE2受体和小细胞外囊泡的相互作用

• 发表时间: 2022
• 作者: Keesiang Lim

High-Speed Atomic Force Microscopy Reveals Spatiotemporal Dynamics of Histone Protein H2A Involution by DNA Inchworming

• DOI: 10.1021/acs.jpclett.1c00697
• 发表时间: 2021-04-14
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY LETTERS
• 影响因子: 5.7
• 作者: Nishide, Goro;Lim, Keesiang;Wong, Richard W.
• 通讯作者: Wong, Richard W.

Conformational dynamics of SARS-CoV-2 spike (S) protein and its interaction with ACE2 receptor and small extracellular vesicles.

SARS-CoV-2 刺突 (S) 蛋白的构象动力学及其与 ACE2 受体和小细胞外囊泡的相互作用。

• 发表时间: 2022
• 作者: Sasaki Eita;Asanuma Hideki;Momose Haruka;Furuhata Keiko;Mizukami Takuo;Matsumura Takayuki;Takahashi Yoshimasa;Hamaguchi Isao;Keesiang Lim
• 通讯作者: Keesiang Lim

High-Speed AFM Reveals Molecular Dynamics of Human Influenza A Hemagglutinin and Its Interaction with Exosomes

• DOI: 10.1021/acs.nanolett.0c01755
• 发表时间: 2020-09-09
• 期刊: NANO LETTERS
• 影响因子: 10.8
• 作者: Lim, Keesiang;Kodera, Noriyuki;Wong, Richard W.
• 通讯作者: Wong, Richard W.