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.

该项目的目标是：1）使用HS-AFM在生理条件下记录天然构象病毒融合蛋白； 2）在使用HS-AFM酸性条件触发的构象变化过程中，可视化快速和短暂的中间体； 3）当使用HS-AFM与外泌体或抗体相互作用时，阐明病毒融合蛋白的构象动态。 PI在与中和抗体的相互作用过程中实施了HS-AFM来对SARS-COV-2尖峰（S）蛋白结构动力学进行纳米镜阐明（目标3）。某些中和抗体促进了RBD从其紧密构象过渡到开放确认，从而增强了RBD-ACE2相互作用。该作用称为抗体依赖性增强作用（ADE）。这项研究表明，HS-AFM可以作为用于S NAB测试的有效纳米镜评估平台。这项研究已发表在良好的期刊《 Nano Letters》中。此外，这项研究也在几个国内会议中提出。登革热病毒的包膜蛋白（E蛋白）将在不久的将来进行研究。

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.