scholarly journals Supported Lipid Bilayer Platform for Characterizing the Membrane-Disruptive Behaviors of Triton X-100 and Potential Detergent Replacements

2022 ◽  
Vol 23 (2) ◽  
pp. 869
Author(s):  
Negin Gooran ◽  
Bo Kyeong Yoon ◽  
Joshua A. Jackman
Keyword(s):  
Lipid Bilayer ◽  
Supported Lipid Bilayer ◽  
Virus Particles ◽  
Membrane Solubilization ◽  
Enveloped Virus ◽  
Interaction Kinetics ◽  
Membrane Budding ◽  
Phase Out ◽  
Triton X ◽  
Interaction Behaviors

Triton X-100 (TX-100) is a widely used detergent to prevent viral contamination of manufactured biologicals and biopharmaceuticals, and acts by disrupting membrane-enveloped virus particles. However, environmental concerns about ecotoxic byproducts are leading to TX-100 phase out and there is an outstanding need to identify functionally equivalent detergents that can potentially replace TX-100. To date, a few detergent candidates have been identified based on viral inactivation studies, while direct mechanistic comparison of TX-100 and potential replacements from a biophysical interaction perspective is warranted. Herein, we employed a supported lipid bilayer (SLB) platform to comparatively evaluate the membrane-disruptive properties of TX-100 and a potential replacement, Simulsol SL 11W (SL-11W), and identified key mechanistic differences in terms of how the two detergents interact with phospholipid membranes. Quartz crystal microbalance-dissipation (QCM-D) measurements revealed that TX-100 was more potent and induced rapid, irreversible, and complete membrane solubilization, whereas SL-11W caused more gradual, reversible membrane budding and did not induce extensive membrane solubilization. The results further demonstrated that TX-100 and SL-11W both exhibit concentration-dependent interaction behaviors and were only active at or above their respective critical micelle concentration (CMC) values. Collectively, our findings demonstrate that TX-100 and SL-11W have distinct membrane-disruptive effects in terms of potency, mechanism of action, and interaction kinetics, and the SLB platform approach can support the development of biophysical assays to efficiently test potential TX-100 replacements.

scholarly journals Conformational Plasticity of Hepatitis B Core Protein Spikes Promotes Peptide Binding Independent of the Secretion Phenotype

2021 ◽  
Vol 9 (5) ◽  
pp. 956
Author(s):  
Cihan Makbul ◽  
Vladimir Khayenko ◽  
Hans Michael Maric ◽  
Bettina Böttcher
Keyword(s):  
Hepatitis B ◽  
Core Protein ◽  
Peptide Binding ◽  
Surface Proteins ◽  
Binding Motif ◽  
Virus Particles ◽  
Enveloped Virus ◽  
Conformational Plasticity ◽  
Viral Maturation ◽  
Hepatitis B Core Protein

Hepatitis B virus is a major human pathogen, which forms enveloped virus particles. During viral maturation, membrane-bound hepatitis B surface proteins package hepatitis B core protein capsids. This process is intercepted by certain peptides with an “LLGRMKG” motif that binds to the capsids at the tips of dimeric spikes. With microcalorimetry, electron cryo microscopy and peptide microarray-based screens, we have characterized the structural and thermodynamic properties of peptide binding to hepatitis B core protein capsids with different secretion phenotypes. The peptide “GSLLGRMKGA” binds weakly to hepatitis B core protein capsids and mutant capsids with a premature (F97L) or low-secretion phenotype (L60V and P5T). With electron cryo microscopy, we provide novel structures for L60V and P5T and demonstrate that binding occurs at the tips of the spikes at the dimer interface, splaying the helices apart independent of the secretion phenotype. Peptide array screening identifies “SLLGRM” as the core binding motif. This shortened motif binds only to one of the two spikes in the asymmetric unit of the capsid and induces a much smaller conformational change. Altogether, these comprehensive studies suggest that the tips of the spikes act as an autonomous binding platform that is unaffected by mutations that affect secretion phenotypes.

Potassium Ion Transport by Valinomycin across a Hg-Supported Lipid Bilayer

2005 ◽  
Vol 127 (38) ◽  
pp. 13316-13323 ◽  
Author(s):  
Lucia Becucci ◽  
Maria Rosa Moncelli ◽  
Renate Naumann ◽  
Rolando Guidelli
Keyword(s):  
Ion Transport ◽  
Lipid Bilayer ◽  
Potassium Ion ◽  
Supported Lipid Bilayer ◽  
Potassium Ion Transport

Characterization of Supported Lipid Bilayer Disruption By Chrysophsin-3 Using QCM-D

2011 ◽  
Vol 115 (51) ◽  
pp. 15228-15235 ◽  
Author(s):  
Kathleen F. Wang ◽  
Ramanathan Nagarajan ◽  
Charlene M. Mello ◽  
Terri A. Camesano
Keyword(s):  
Lipid Bilayer ◽  
Supported Lipid Bilayer

Formation of Supported Lipid Bilayer Membranes on SiO2from Proteoliposomes Containing Transmembrane Proteins

Langmuir ◽  
2003 ◽  
Vol 19 (3) ◽  
pp. 842-850 ◽  
Author(s):  
Annette Granéli ◽  
Jan Rydström ◽  
Bengt Kasemo ◽  
Fredrik Höök
Keyword(s):  
Lipid Bilayer ◽  
Transmembrane Proteins ◽  
Lipid Bilayer Membranes ◽  
Supported Lipid Bilayer ◽  
Bilayer Membranes

A hybrid SAM phospholipid approach to fabricating a ‘free’ supported lipid bilayer

2002 ◽  
Vol 4 (11) ◽  
pp. 2371-2378 ◽  
Author(s):  
Arwel V. Hughes ◽  
Arach Goldar ◽  
Michael C. Gerstenberg ◽  
Steve J. Roser ◽  
Jeremy Bradshaw
Keyword(s):  
Lipid Bilayer ◽  
Supported Lipid Bilayer

scholarly journals A non-enveloped arbovirus released in lysosome-derived extracellular vesicles induces super-infection exclusion

2020 ◽  
Author(s):  
Thomas Labadie ◽  
Polly Roy
Keyword(s):  
Extracellular Vesicles ◽  
Defense Mechanism ◽  
Degradation Process ◽  
Free Virus ◽  
Enveloped Viruses ◽  
Virus Particles ◽  
Enveloped Virus ◽  
Super Infection

AbstractRecent developments on extracellular vesicles (EVs) containing multiple virus particles challenge the rigid definition of non-enveloped viruses. However, how non-enveloped viruses hijack cell machinery to promote non-lytic release in EVs, and their functional roles, remain to be clarified. Here we used Bluetongue virus (BTV) as a model of a non-enveloped arthropod-borne virus and observed that the majority of viruses are released in EVs, both in vitro and in the blood of infected animals. Based on the cellular proteins detected in these EVs, and use of inhibitors targeting the cellular degradation process, we demonstrated that these extracellular vesicles are derived from secretory lysosomes, in which the acidic pH is neutralized upon the infection. Moreover, we report that secreted EVs are more efficient than free-viruses for initiating infections, but that they trigger super-infection exclusion that only free-viruses can overcome.Author summaryRecent discoveries of non-enveloped virus secreted in EVs opened the door to new developments in our understanding of the transmission and pathogenicity of these viruses. In particular, how these viruses hijack the host cellular secretion machinery, and the role of these EVs compared with free-virus particles remained to be explored. Here, we tackled these two aspects, by studying BTV, an emerging arthropod-borne virus causing epidemics worldwide. We showed that this virus is mainly released in EVs, in vivo and in the blood of infected animals, and that inhibition of the cell degradation machinery decreases the release of infectious EVs, but not free-virus particles. We found that BTV must neutralize the pH of lysosomes, which are important organelles of the cell degradation machinery, for efficient virus release in EVs. Our results highlight unique features for a virus released in EVs, explaining how BTV transits in lysosomes without being degraded. Interestingly, we observed that EVs are more infectious than free-virus particles, but only free-viruses are able to overcome the super-infection exclusion, which is a common cellular defense mechanism. In conclusion, our study stresses the dual role played by both forms, free and vesicular, in the virus life cycle.

scholarly journals The vaccinia virus F12L protein is associated with intracellular enveloped virus particles and is required for their egress to the cell surface

2002 ◽  
Vol 83 (1) ◽  
pp. 195-207 ◽  
Author(s):  
Henriette van Eijl ◽  
Michael Hollinshead ◽  
Gaener Rodger ◽  
Wei-Hong Zhang ◽  
Geoffrey L. Smith
Keyword(s):  
Cell Surface ◽  
Vaccinia Virus ◽  
Virus Particles ◽  
Enveloped Virus ◽  
Virus Morphogenesis ◽  
C Terminus ◽  
Specific Proteins ◽  
Infected Cells ◽  
Mature Virus ◽  
Confocal And Electron Microscopy

The vaccinia virus (VV) F12L gene encodes a 65 kDa protein that is expressed late during infection and is important for plaque formation, EEV production and virulence. Here we have used a recombinant virus (vF12LHA) in which the F12L protein is tagged at the C terminus with an epitope recognized by a monoclonal antibody to determine the location of F12L in infected cells and whether it associates with virions. Using confocal and electron microscopy we show that the F12L protein is located on intracellular enveloped virus (IEV) particles, but is absent from immature virions (IV), intracellular mature virus (IMV) and cell-associated enveloped virus (CEV). In addition, F12L shows co-localization with endosomal compartments and microtubules. F12L did not co-localize with virions attached to actin tails, providing further evidence that actin tails are associated with CEV but not IEV particles. In vΔF12L-infected cells, virus morphogenesis was arrested after the formation of IEV particles, so that the movement of these virions to the cell surface was inhibited and CEV particles were not found. Previously, virus mutants lacking IEV- or EEV-specific proteins were either unable to make IEV particles (vΔF13L and vΔB5R), or were unable to form actin tails after formation of CEV particles (vΔA36R, vΔA33R, vΔA34R). The F12L deletion mutant therefore defines a new stage in the morphogenic pathway and the F12L protein is implicated as necessary for microtubule-mediated egress of IEV particles to the cell surface.

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