[PMID]: | 29095961 |
[Au] Autor: | Koromyslova AD; Hansman GS |
[Ad] Endereço: | Schaller Research Group at the University of Heidelberg and the DKFZ, Heidelberg, Germany. |
[Ti] Título: | Nanobodies targeting norovirus capsid reveal functional epitopes and potential mechanisms of neutralization. |
[So] Source: | PLoS Pathog;13(11):e1006636, 2017 Nov. |
[Is] ISSN: | 1553-7374 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs) are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important. In the current study, we investigated a panel of Nanobodies in order to probe functional epitopes that could trigger capsid rearrangement and/ or interfere with HBGA binding interactions. The precise binding sites of six Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) were identified using X-ray crystallography. We showed that these Nanobodies bound on the top, side, and bottom of the norovirus protruding domain. The impact of Nanobody binding on norovirus capsid morphology was analyzed using electron microscopy and dynamic light scattering. We discovered that distinct Nanobody epitopes were associated with varied changes in particle structural integrity and assembly. Interestingly, certain Nanobody-induced capsid morphological changes lead to the capsid protein degradation and viral RNA exposure. Moreover, Nanobodies employed multiple inhibition mechanisms to prevent norovirus attachment to HBGAs, which included steric obstruction (Nano-14), allosteric interference (Nano-32), and violation of normal capsid morphology (Nano-26 and Nano-85). Finally, we showed that two Nanobodies (Nano-26 and Nano-85) not only compromised capsid integrity and inhibited VLPs attachment to HBGAs, but also recognized a broad panel of norovirus genotypes with high affinities. Consequently, Nano-26 and Nano-85 have a great potential to function as novel therapeutic agents against human noroviruses. |
[Mh] Termos MeSH primário: |
Anticorpos Neutralizantes/farmacologia Antivirais/farmacologia Proteínas do Capsídeo/antagonistas & inibidores Capsídeo/efeitos dos fármacos Modelos Moleculares Norovirus/efeitos dos fármacos Anticorpos de Domínio Único/farmacologia
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[Mh] Termos MeSH secundário: |
Anticorpos Neutralizantes/química Anticorpos Neutralizantes/metabolismo Afinidade de Anticorpos Antivirais/química Antivirais/metabolismo Sítios de Ligação de Anticorpos Ligação Competitiva Antígenos de Grupos Sanguíneos/química Antígenos de Grupos Sanguíneos/metabolismo Capsídeo/química Capsídeo/metabolismo Capsídeo/ultraestrutura Proteínas do Capsídeo/química Proteínas do Capsídeo/metabolismo Reações Cruzadas Cristalografia por Raios X Difusão Dinâmica da Luz Epitopos Cinética Microscopia Eletrônica de Transmissão Norovirus/química Norovirus/metabolismo Norovirus/ultraestrutura Conformação Proteica Domínios e Motivos de Interação entre Proteínas Anticorpos de Domínio Único/química Anticorpos de Domínio Único/metabolismo Termodinâmica
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[Pt] Tipo de publicação: | COMPARATIVE STUDY; JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Antibodies, Neutralizing); 0 (Antiviral Agents); 0 (Blood Group Antigens); 0 (Capsid Proteins); 0 (Epitopes); 0 (Single-Domain Antibodies) |
[Em] Mês de entrada: | 1711 |
[Cu] Atualização por classe: | 171119 |
[Lr] Data última revisão:
| 171119 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 171103 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1371/journal.ppat.1006636 |
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