2020 Fiscal Year Research-status Report
Real-time observation of native conformations and molecular behaviors of viral fusion proteins using high-speed atomic force microscopy (HS-AFM)
| Project/Area Number |
20K16262
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| Research Institution | Kanazawa University |
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Principal Investigator |
LIM KEE・SIANG 金沢大学, ナノ生命科学研究所, 特任助教 (60842987)
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Keywords | Viral fusion protein / fusogenic transition / exosome / HS-AFM |
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| Outline of Annual Research Achievements |
In summary, this project is running smoothly, and all aims have been completed for the influenza A hemagglutinin (HA). The native conformation of human influenza A hemagglutinin (HA) observed using HS-AFM is ellipsoidal, and it undergoes fusogenic transition in acidic condition to become Y-shape. Direct real-time observation of fusogenic transition suggests that the transition mechanism is likely to fit to the Uncaging Model. HA-exosome interaction is weak in neutral condition but firm in acidic condition. The weak interaction could be mediated by HA-receptor (sialic acid) interaction. In contrast, the firm interaction implies that HA fusion peptide could be released after fusogenic transition, and then inserted into exosomal layer and resulted deformation or rupture of exosomes.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Three aims have been completed for one viral fusion protein, the influenza A virus hemagglutinin, and published in Nano Letters. Four different viral proteins are proposed for this study including influenza A virus hemagglutinin, dengue virus E protein, Zika virus E protein, and Rabies virus G protein. With this pace, to complete the remaining three viral fusion proteins within the grant period is possible.
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| Strategy for Future Research Activity |
The coming research direction will be focused on Dengue E protein. Experiments will be conducted on Dengue E protein to fulfill the three aims proposed in the grant proposal. Furthermore, Dengue virus-like particle, which is safe for BSL1 experiment, will also be purchased to mimic real viral morphology for Hs-AFM observation.
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| Causes of Carryover |
Research grant will be used for purchasing Dengue virus-related products and HS-AFM consumables. Dengue virus-related products including recombinant Dengue E proteins (several serotypes), Dengue E protein antibodies, and Dengue virus-like particles. HS-AFM consumables including mica, glass stage, and cantilevers. These materials will be used for studying conformational dynamic of Dengue E proteins using HS-AFM.
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