Developments in single-molecule and single-particle fluorescence-based approaches for studying viral envelope glycoprotein dynamics and membrane fusion

Howard, Angela R.; Munro, James B.

doi:10.1016/bs.aivir.2019.05.004

Cited by 5 publications

(6 citation statements)

References 95 publications

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“…CD4-binding induces conformational changes in gp120 to expose coreceptor-binding sites, whereas CCR5 or CXCR4 binding to gp120 is thought to activate refolding events in gp41, eventually leading to viral membrane fusion. smFRET has been used to characterise the conformational changes in gp120 during target receptor binding, the findings of which have been summarised in a number of excellent reviews (Howard and Munro, 2019;Lu et al, 2019b;Lu, 2021). Munro et al (2014) assessed the movement of variable loops in gp120 during receptor binding by labelling the V1 loop with a donor fluorophore, and the V4 or V5 loops with an acceptor fluorophore, followed by smFRET analysis via TIRF microscopy (Figure 1).…”

Section: The Hiv-1 Envelope Proteinmentioning

confidence: 99%

Single-molecule FRET for virology: 20 years of insight into protein structure and dynamics

Groves¹,

Hepp²,

Kapanidis³

et al. 2023

Quart. Rev. Biophys.

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Although viral protein structure and replication mechanisms have been explored extensively with X-ray crystallography, cryo-electron microscopy, and population imaging studies, these methods are often not able to distinguish dynamic conformational changes in real time. Single-molecule fluorescence resonance energy transfer (smFRET) offers unique insights into interactions and states that may be missed in ensemble studies, such as nucleic acid or protein structure, and conformational transitions during folding, receptor–ligand interactions, and fusion. We discuss the application of smFRET to the study of viral protein conformational dynamics, with a particular focus on viral glycoprotein dynamics, viral helicases, proteins involved in HIV reverse transcription, and the influenza RNA polymerase. smFRET experiments have played a crucial role in deciphering conformational changes in these processes, emphasising the importance of smFRET as a tool to help elucidate the life cycle of viral pathogens and identify key anti-viral targets.

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Section: The Hiv-1 Envelope Proteinmentioning

confidence: 99%

Single-molecule FRET for virology: 20 years of insight into protein structure and dynamics

Groves¹,

Hepp²,

Kapanidis³

et al. 2023

Quart. Rev. Biophys.

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“…Imaging macromolecules at the single-molecule/single-particle level has advanced our understanding of both static and dynamic aspects in virus–host interactions, merited by avoiding the averaging-out effect from traditional ensemble-level measurements. Those imaging techniques, such as single-particle cryoEM/cryoET, single-particle optical tracking, and super-resolution fluorescence microscopy exerted significant roles in addressing fundamental questions with regards to structures, dynamics, and functions of virus molecules underlying virus–host interactions [ 38 , 39 , 40 , 41 , 42 , 43 , 44 ].…”

Section: Single-molecule Förster Resonance Energy Transfer (Smfretmentioning

confidence: 99%

“…One unique strength of smFRET is to reveal the timing, order, and frequency of conformational states and state-to-state transitions of S in situ on the surface of viral particles ( Figure 3 D) [ 40 , 41 , 47 ]. Kinetic analyses deployed in smFRET include the transition density for state-to-state transition (plotted as initial state vs. final state), the hidden Markov modeling for idealizing molecular motions, and dwelling time for estimating transitional rates.…”

Section: Conformational Dynamics Of Virus Spike Proteins On the Sumentioning

confidence: 99%

Single-Molecule FRET Imaging of Virus Spike–Host Interactions

2021

Viruses

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As a major surface glycoprotein of enveloped viruses, the virus spike protein is a primary target for vaccines and anti-viral treatments. Current vaccines aiming at controlling the COVID-19 pandemic are mostly directed against the SARS-CoV-2 spike protein. To promote virus entry and facilitate immune evasion, spikes must be dynamic. Interactions with host receptors and coreceptors trigger a cascade of conformational changes/structural rearrangements in spikes, which bring virus and host membranes in proximity for membrane fusion required for virus entry. Spike-mediated viral membrane fusion is a dynamic, multi-step process, and understanding the structure–function-dynamics paradigm of virus spikes is essential to elucidate viral membrane fusion, with the ultimate goal of interventions. However, our understanding of this process primarily relies on individual structural snapshots of endpoints. How these endpoints are connected in a time-resolved manner, and the order and frequency of conformational events underlying virus entry, remain largely elusive. Single-molecule Förster resonance energy transfer (smFRET) has provided a powerful platform to connect structure–function in motion, revealing dynamic aspects of spikes for several viruses: SARS-CoV-2, HIV-1, influenza, and Ebola. This review focuses on how smFRET imaging has advanced our understanding of virus spikes’ dynamic nature, receptor-binding events, and mechanism of antibody neutralization, thereby informing therapeutic interventions.

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“…Single-molecule fluorescence resonance energy transfer experiments indicate that, after binding to a single CD4 molecule, the individual protomers have distinct conformations. The trimer undergoes further transformation when the remaining gp120 molecules bind CD4 and the co-receptor [15,16].…”

Section: Hiv-1 21 Hiv-1 Infection Is Mediated By the Viral Envelope G...mentioning

confidence: 99%

Inhibition of Viral Membrane Fusion by Peptides and Approaches to Peptide Design

2021

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Fusion of lipid-enveloped viruses with the cellular plasma membrane or the endosome membrane is mediated by viral envelope proteins that undergo large conformational changes following binding to receptors. The HIV-1 fusion protein gp41 undergoes a transition into a “six-helix bundle” after binding of the surface protein gp120 to the CD4 receptor and a co-receptor. Synthetic peptides that mimic part of this structure interfere with the formation of the helix structure and inhibit membrane fusion. This approach also works with the S spike protein of SARS-CoV-2. Here we review the peptide inhibitors of membrane fusion involved in infection by influenza virus, HIV-1, MERS and SARS coronaviruses, hepatitis viruses, paramyxoviruses, flaviviruses, herpesviruses and filoviruses. We also describe recent computational methods used for the identification of peptide sequences that can interact strongly with protein interfaces, with special emphasis on SARS-CoV-2, using the PePI-Covid19 database.

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Developments in single-molecule and single-particle fluorescence-based approaches for studying viral envelope glycoprotein dynamics and membrane fusion

Cited by 5 publications

References 95 publications

Single-molecule FRET for virology: 20 years of insight into protein structure and dynamics

Single-molecule FRET for virology: 20 years of insight into protein structure and dynamics

Single-Molecule FRET Imaging of Virus Spike–Host Interactions

Inhibition of Viral Membrane Fusion by Peptides and Approaches to Peptide Design

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