A Vulnerable, Membrane-Proximal Site in Human Respiratory Syncytial Virus F Revealed by a Prefusion-Specific Single-Domain Antibody

Rossey, Iebe; Hsieh, Ching Lin; Sedeyn, Koen; Ballegeer, Marlies; Schepens, Bert; McLellan, Jason S.; Saelens, Xavier

doi:10.1128/jvi.02279-20

Cited by 9 publications

(6 citation statements)

References 33 publications

(50 reference statements)

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“…Lastly, designs consisting of linear fusion proteins do not allow the inclusion of binders that require a free N-terminus for target recognition. 17 …”

Section: Resultsmentioning

confidence: 99%

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

Gräwe,

Spruit,

de Vries

et al. 2024

RSC Chem. Biol.

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“…Lastly, designs consisting of linear fusion proteins do not allow the inclusion of binders that require a free N-terminus for target recognition. 17 …”

Section: Resultsmentioning

confidence: 99%

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

Gräwe,

Spruit,

de Vries

et al. 2024

RSC Chem. Biol.

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“…Moreover, the integration of multiple binder domains for viral protein targets (instead of simple peptide epitopes) would result in complex, multidomain fusion proteins whose expression could prove difficult due to misfolding during translation. Lastly, designs consisting of linear fusion proteins do not allow the inclusion of binders that require a free N-terminus for target recognition 17 .…”

Section: Resultsmentioning

confidence: 99%

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

Gräwe,

Spruit,

de Vries

et al. 2023

Preprint

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Fast and reliable virus diagnostics is key to prevent the spread of viruses in populations. A hallmark of viruses is the presence of multivalent surface proteins, a property that can be harnessed to control conformational switching in sensor proteins. Here, we introduce a new sensor platform (dark-LUX) for the detection of viral surface proteins consisting of a general bioluminescent framework that can be post-translationally functionalized with separately expressed binding domains. The platform relies on 1) plug-and-play bioconjugation of different binding proteins via SpyTag/SpyCatcher technology to create branched protein structures, 2) an optimized turn-on bioluminescent switch based on complementation of the split-luciferase NanoBiT upon target binding and 3) straightforward exploration of the protein linker space. The influenza A virus (IAV) surface proteins hemagglutinin (HA) and neuraminidase (NA) were used as relevant multivalent targets to establish proof of principle and optimize relevant parameters such as linker properties, choice of target binding domains and the optimal combination of the competing NanoBiT components SmBiT and DarkBiT. The sensor framework allows rapid conjugation and exchange of various binding domains including scFvs, nanobodies and de novo designed binders for a variety of targets, including the construction of a heterobivalent switch that targets the head and stem region of hemagglutinin. The modularity of the platform thus allows straightforward optimization of binding domains and scaffold properties for existing viral targets, and is well suited to quickly adapt bioluminescent sensor proteins to effectively detect newly evolving viral epitopes.

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“…The geometrical parameters of full-size F protein (PDB code 7LVW [ 25 ] of human RVS A2 were downloaded from non-commercial database Protein Data Bank [ 26 ]. To localization of the binding site complex protomer, inhibitor RV521 was used (PDB code 7KQD [ 7 ].…”

Section: Methodsmentioning

confidence: 99%

New Inhibitors of Respiratory Syncytial Virus (RSV) Replication Based on Monoterpene-Substituted Arylcoumarins

et al. 2023

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Respiratory syncytial virus (RSV) causes annual epidemics of respiratory infection. Usually harmless to adults, the RSV infection can be dangerous to children under 3 years of age and elderly people over 65 years of age, often causing serious problems, even death. At present, there are no vaccines and specific chemotherapeutic agents for the treatment of this disease, so the search for low-molecular weight compounds to combat RSV is a challenge. In this work, we have shown, for the first time, that monoterpene-substituted arylcoumarins are efficient RSV replication inhibitors at low micromolar concentrations. The most active compound has a selectivity index of about 200 and acts most effectively at the early stages of infection. The F protein of RSV is a potential target for these compounds, which is also confirmed by molecular docking and molecular dynamics simulation data.

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A Vulnerable, Membrane-Proximal Site in Human Respiratory Syncytial Virus F Revealed by a Prefusion-Specific Single-Domain Antibody

Cited by 9 publications

References 33 publications

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

Bioluminescent detection of viral surface proteins using branched multivalent protein switches

New Inhibitors of Respiratory Syncytial Virus (RSV) Replication Based on Monoterpene-Substituted Arylcoumarins

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