A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus.
Recombinant subunit vaccines should contain minimal non-pathogen motifs to reduce potential off-target reactivity. We recently developed a vaccine antigen against respiratory syncytial virus (RSV), which comprised the fusion (F) glycoprotein stabilized in its pre-fusion trimeric conformation by &quo...
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doaj-842b18df124241c789b75fea7778ce7c2020-11-25T02:13:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01106e012877910.1371/journal.pone.0128779A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus.Guillaume B E Stewart-JonesPaul V ThomasMan ChenAliaksandr DruzM Gordon JoyceWing-Pui KongMallika SastryCinque SotoYongping YangBaoshan ZhangLei ChenGwo-Yu ChuangIvelin S GeorgievJason S McLellanSanjay SrivatsanTongqing ZhouUlrich BaxaJohn R MascolaBarney S GrahamPeter D KwongRecombinant subunit vaccines should contain minimal non-pathogen motifs to reduce potential off-target reactivity. We recently developed a vaccine antigen against respiratory syncytial virus (RSV), which comprised the fusion (F) glycoprotein stabilized in its pre-fusion trimeric conformation by "DS-Cav1" mutations and by an appended C-terminal trimerization motif or "foldon" from T4-bacteriophage fibritin. Here we investigate the creation of a cysteine zipper to allow for the removal of the phage foldon, while maintaining the immunogenicity of the parent DS-Cav1+foldon antigen. Constructs without foldon yielded RSV F monomers, and enzymatic removal of the phage foldon from pre-fusion F trimers resulted in their dissociation into monomers. Because the native C terminus of the pre-fusion RSV F ectodomain encompasses a viral trimeric coiled-coil, we explored whether introduction of cysteine residues capable of forming inter-protomer disulfides might allow for stable trimers. Structural modeling indicated the introduced cysteines to form disulfide "rings", with each ring comprising a different set of inward facing residues of the coiled-coil. Three sets of rings could be placed within the native RSV F coiled-coil, and additional rings could be added by duplicating portions of the coiled-coil. High levels of neutralizing activity in mice, equivalent to that of the parent DS-Cav1+foldon antigen, were elicited by a 4-ring stabilized RSV F trimer with no foldon. Structure-based alteration of a viral coiled-coil to create a cysteine zipper thus allows a phage trimerization motif to be removed from a candidate vaccine antigen.http://europepmc.org/articles/PMC4476739?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Guillaume B E Stewart-Jones Paul V Thomas Man Chen Aliaksandr Druz M Gordon Joyce Wing-Pui Kong Mallika Sastry Cinque Soto Yongping Yang Baoshan Zhang Lei Chen Gwo-Yu Chuang Ivelin S Georgiev Jason S McLellan Sanjay Srivatsan Tongqing Zhou Ulrich Baxa John R Mascola Barney S Graham Peter D Kwong |
spellingShingle |
Guillaume B E Stewart-Jones Paul V Thomas Man Chen Aliaksandr Druz M Gordon Joyce Wing-Pui Kong Mallika Sastry Cinque Soto Yongping Yang Baoshan Zhang Lei Chen Gwo-Yu Chuang Ivelin S Georgiev Jason S McLellan Sanjay Srivatsan Tongqing Zhou Ulrich Baxa John R Mascola Barney S Graham Peter D Kwong A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. PLoS ONE |
author_facet |
Guillaume B E Stewart-Jones Paul V Thomas Man Chen Aliaksandr Druz M Gordon Joyce Wing-Pui Kong Mallika Sastry Cinque Soto Yongping Yang Baoshan Zhang Lei Chen Gwo-Yu Chuang Ivelin S Georgiev Jason S McLellan Sanjay Srivatsan Tongqing Zhou Ulrich Baxa John R Mascola Barney S Graham Peter D Kwong |
author_sort |
Guillaume B E Stewart-Jones |
title |
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. |
title_short |
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. |
title_full |
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. |
title_fullStr |
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. |
title_full_unstemmed |
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus. |
title_sort |
cysteine zipper stabilizes a pre-fusion f glycoprotein vaccine for respiratory syncytial virus. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2015-01-01 |
description |
Recombinant subunit vaccines should contain minimal non-pathogen motifs to reduce potential off-target reactivity. We recently developed a vaccine antigen against respiratory syncytial virus (RSV), which comprised the fusion (F) glycoprotein stabilized in its pre-fusion trimeric conformation by "DS-Cav1" mutations and by an appended C-terminal trimerization motif or "foldon" from T4-bacteriophage fibritin. Here we investigate the creation of a cysteine zipper to allow for the removal of the phage foldon, while maintaining the immunogenicity of the parent DS-Cav1+foldon antigen. Constructs without foldon yielded RSV F monomers, and enzymatic removal of the phage foldon from pre-fusion F trimers resulted in their dissociation into monomers. Because the native C terminus of the pre-fusion RSV F ectodomain encompasses a viral trimeric coiled-coil, we explored whether introduction of cysteine residues capable of forming inter-protomer disulfides might allow for stable trimers. Structural modeling indicated the introduced cysteines to form disulfide "rings", with each ring comprising a different set of inward facing residues of the coiled-coil. Three sets of rings could be placed within the native RSV F coiled-coil, and additional rings could be added by duplicating portions of the coiled-coil. High levels of neutralizing activity in mice, equivalent to that of the parent DS-Cav1+foldon antigen, were elicited by a 4-ring stabilized RSV F trimer with no foldon. Structure-based alteration of a viral coiled-coil to create a cysteine zipper thus allows a phage trimerization motif to be removed from a candidate vaccine antigen. |
url |
http://europepmc.org/articles/PMC4476739?pdf=render |
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