Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus.
We currently have an incomplete understanding of why only a fraction of human antibodies that bind to flaviviruses block infection of cells. Here we define the footprint of a strongly neutralizing human monoclonal antibody (mAb G9E) with Zika virus (ZIKV) by both X-ray crystallography and cryo-elect...
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Public Library of Science (PLoS)
2023-01-01
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| Series: | PLoS Pathogens |
| Online Access: | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010814&type=printable |
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| author | Cameron Adams Derek L Carbaugh Bo Shu Thiam-Seng Ng Izabella N Castillo Ryan Bhowmik Bruno Segovia-Chumbez Ana C Puhl Stephen Graham Sean A Diehl Helen M Lazear Shee-Mei Lok Aravinda M de Silva Lakshmanane Premkumar |
| author_facet | Cameron Adams Derek L Carbaugh Bo Shu Thiam-Seng Ng Izabella N Castillo Ryan Bhowmik Bruno Segovia-Chumbez Ana C Puhl Stephen Graham Sean A Diehl Helen M Lazear Shee-Mei Lok Aravinda M de Silva Lakshmanane Premkumar |
| author_sort | Cameron Adams |
| collection | DOAJ |
| description | We currently have an incomplete understanding of why only a fraction of human antibodies that bind to flaviviruses block infection of cells. Here we define the footprint of a strongly neutralizing human monoclonal antibody (mAb G9E) with Zika virus (ZIKV) by both X-ray crystallography and cryo-electron microscopy. Flavivirus envelope (E) glycoproteins are present as homodimers on the virion surface, and G9E bound to a quaternary structure epitope spanning both E protomers forming a homodimer. As G9E mainly neutralized ZIKV by blocking a step after viral attachment to cells, we tested if the neutralization mechanism of G9E was dependent on the mAb cross-linking E molecules and blocking low-pH triggered conformational changes required for viral membrane fusion. We introduced targeted mutations to the G9E paratope to create recombinant antibodies that bound to the ZIKV envelope without cross-linking E protomers. The G9E paratope mutants that bound to a restricted epitope on one protomer poorly neutralized ZIKV compared to the wild-type mAb, demonstrating that the neutralization mechanism depended on the ability of G9E to cross-link E proteins. In cell-free low pH triggered viral fusion assay, both wild-type G9E, and epitope restricted paratope mutant G9E bound to ZIKV but only the wild-type G9E blocked fusion. We propose that, beyond antibody binding strength, the ability of human antibodies to cross-link E-proteins is a critical determinant of flavivirus neutralization potency. |
| format | Article |
| id | doaj-art-470ce3319ca643cab15d968be35cdd93 |
| institution | DOAJ |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-470ce3319ca643cab15d968be35cdd932025-08-20T03:08:32ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742023-01-01191e101081410.1371/journal.ppat.1010814Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus.Cameron AdamsDerek L CarbaughBo ShuThiam-Seng NgIzabella N CastilloRyan BhowmikBruno Segovia-ChumbezAna C PuhlStephen GrahamSean A DiehlHelen M LazearShee-Mei LokAravinda M de SilvaLakshmanane PremkumarWe currently have an incomplete understanding of why only a fraction of human antibodies that bind to flaviviruses block infection of cells. Here we define the footprint of a strongly neutralizing human monoclonal antibody (mAb G9E) with Zika virus (ZIKV) by both X-ray crystallography and cryo-electron microscopy. Flavivirus envelope (E) glycoproteins are present as homodimers on the virion surface, and G9E bound to a quaternary structure epitope spanning both E protomers forming a homodimer. As G9E mainly neutralized ZIKV by blocking a step after viral attachment to cells, we tested if the neutralization mechanism of G9E was dependent on the mAb cross-linking E molecules and blocking low-pH triggered conformational changes required for viral membrane fusion. We introduced targeted mutations to the G9E paratope to create recombinant antibodies that bound to the ZIKV envelope without cross-linking E protomers. The G9E paratope mutants that bound to a restricted epitope on one protomer poorly neutralized ZIKV compared to the wild-type mAb, demonstrating that the neutralization mechanism depended on the ability of G9E to cross-link E proteins. In cell-free low pH triggered viral fusion assay, both wild-type G9E, and epitope restricted paratope mutant G9E bound to ZIKV but only the wild-type G9E blocked fusion. We propose that, beyond antibody binding strength, the ability of human antibodies to cross-link E-proteins is a critical determinant of flavivirus neutralization potency.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010814&type=printable |
| spellingShingle | Cameron Adams Derek L Carbaugh Bo Shu Thiam-Seng Ng Izabella N Castillo Ryan Bhowmik Bruno Segovia-Chumbez Ana C Puhl Stephen Graham Sean A Diehl Helen M Lazear Shee-Mei Lok Aravinda M de Silva Lakshmanane Premkumar Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. PLoS Pathogens |
| title | Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. |
| title_full | Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. |
| title_fullStr | Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. |
| title_full_unstemmed | Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. |
| title_short | Structure and neutralization mechanism of a human antibody targeting a complex Epitope on Zika virus. |
| title_sort | structure and neutralization mechanism of a human antibody targeting a complex epitope on zika virus |
| url | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010814&type=printable |
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