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      Probing structural changes in single enveloped virus particles using nano-infrared spectroscopic imaging

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          Abstract

          Enveloped viruses, such as HIV, Ebola and Influenza, are among the most deadly known viruses. Cellular membrane penetration of enveloped viruses is a critical step in the cascade of events that lead to entry into the host cell. Conventional ensemble fusion assays rely on collective responses to membrane fusion events, and do not allow direct and quantitative studies of the subtle and intricate fusion details. Such details are accessible via single particle investigation techniques, however. Here, we implement nano-infrared spectroscopic imaging to investigate the chemical and structural modifications that occur prior to membrane fusion in the single archetypal enveloped virus, influenza X31. We traced in real-space structural and spectroscopic alterations that occur during environmental pH variations in single virus particles. In addition, using nanospectroscopic imaging we quantified the effectiveness of an antiviral compound in stopping viral membrane disruption (a novel mechanism for inhibiting viral entry into cells) during environmental pH variations.

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          Most cited references27

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          Nano-FTIR absorption spectroscopy of molecular fingerprints at 20 nm spatial resolution.

          We demonstrate Fourier transform infrared nanospectroscopy (nano-FTIR) based on a scattering-type scanning near-field optical microscope (s-SNOM) equipped with a coherent-continuum infrared light source. We show that the method can straightforwardly determine the infrared absorption spectrum of organic samples with a spatial resolution of 20 nm, corresponding to a probed volume as small as 10 zeptoliter (10(-20) L). Corroborated by theory, the nano-FTIR absorption spectra correlate well with conventional FTIR absorption spectra, as experimentally demonstrated with poly(methyl methacrylate) (PMMA) samples. Nano-FTIR can thus make use of standard infrared databases of molecular vibrations to identify organic materials in ultrasmall quantities and at ultrahigh spatial resolution. As an application example we demonstrate the identification of a nanoscale PDMS contamination on a PMMA sample.
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            Virus and cell fusion mechanisms.

            In biomembrane fusion pathways, membranes are destabilized through insertions of amphipathic protein segments, lipid reorganization via hemifusion, protein restructuring, and dimpling of the membranes. Four classes of membrane proteins are known in virus and cell fusion. Class I virus-cell fusion proteins (fusogens) are α-helix-rich prefusion trimers that form coiled-coil structures that insert hydrophobic fusion peptides or loops (FPs or FLs) into membranes and refold into postfusion trimers. Class II virus-cell fusogens are β-sheet-rich prefusion homo- or heterodimers that insert FLs into membranes, ending in postfusion trimers. Class III virus-cell fusogens are trimers with both α-helices and β-sheets that dissociate into monomers, insert FLs into membranes, and oligomerize into postfusion trimers. Class IV reoviral cell-cell fusogens are small proteins with FLs that oligomerize to fuse membranes. Class I cell-cell fusogens (Syncytins) were captured by mammals from retroviruses, and class II cell-cell fusogens (EFF-1/AFF-1) fuse membranes via homotypic zippering. Mechanisms and fusogens for most cell fusion events are unknown.
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              Nano-optical imaging and spectroscopy of order, phases, and domains in complex solids

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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Writing – review & editing
                Role: Formal analysis
                Role: Data curationRole: Formal analysis
                Role: Data curationRole: Formal analysis
                Role: Data curationRole: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                12 June 2018
                2018
                : 13
                : 6
                : e0199112
                Affiliations
                [1 ] Department of Physics and Astronomy, University of Georgia, Athens, Georgia, United States of America
                [2 ] Department of Physics and Astronomy, Georgia State University, Atlanta, Georgia, United States of America
                [3 ] Department of Chemistry, Georgia State University, Atlanta, Georgia, United States of America
                Centre de Recherche en Cancerologie de Lyon, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0003-2044-0698
                http://orcid.org/0000-0002-9299-8693
                Article
                PONE-D-18-09566
                10.1371/journal.pone.0199112
                5997350
                29894493
                2776dd60-8f0b-4150-b87b-05a74f52f049
                © 2018 Gamage et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 30 March 2018
                : 31 May 2018
                Page count
                Figures: 6, Tables: 0, Pages: 12
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: 1553251
                Award Recipient :
                This work was supported by the National Science Foundation, Award Number: 1553251, ( https://www.nsf.gov/) to YA. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Microbiology
                Virology
                Viral Structure
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Cell Membranes
                Membrane Fusion
                Biology and life sciences
                Organisms
                Viruses
                RNA viruses
                Orthomyxoviruses
                Influenza Viruses
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Viral Pathogens
                Orthomyxoviruses
                Influenza Viruses
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Viral Pathogens
                Orthomyxoviruses
                Influenza Viruses
                Biology and Life Sciences
                Organisms
                Viruses
                Viral Pathogens
                Orthomyxoviruses
                Influenza Viruses
                Biology and Life Sciences
                Microbiology
                Virology
                Viral Transmission and Infection
                Viral Entry
                Research and Analysis Methods
                Spectrum Analysis Techniques
                Absorption Spectroscopy
                Biology and Life Sciences
                Biochemistry
                Lipids
                Physical Sciences
                Chemistry
                Chemical Compounds
                Organic Compounds
                Amides
                Physical Sciences
                Chemistry
                Organic Chemistry
                Organic Compounds
                Amides
                Biology and Life Sciences
                Microbiology
                Virology
                Viral Transmission and Infection
                Host Cells
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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