Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo.
Enveloped viruses utilize membrane glycoproteins on their surface to mediate entry into host cells. Three-dimensional structural analysis of these glycoprotein 'spikes' is often technically challenging but important for understanding viral pathogenesis and in drug design. Here, a protocol...
| Hoofdauteurs: | , , , , , |
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| Formaat: | Journal article |
| Taal: | English |
| Gepubliceerd in: |
Journal of Visualized Experiments
2014
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| _version_ | 1826298587591475200 |
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| author | Huiskonen, J Parsy, M Li, S Bitto, D Renner, M Bowden, T |
| author_facet | Huiskonen, J Parsy, M Li, S Bitto, D Renner, M Bowden, T |
| author_sort | Huiskonen, J |
| collection | OXFORD |
| description | Enveloped viruses utilize membrane glycoproteins on their surface to mediate entry into host cells. Three-dimensional structural analysis of these glycoprotein 'spikes' is often technically challenging but important for understanding viral pathogenesis and in drug design. Here, a protocol is presented for viral spike structure determination through computational averaging of electron cryo-tomography data. Electron cryo-tomography is a technique in electron microscopy used to derive three-dimensional tomographic volume reconstructions, or tomograms, of pleomorphic biological specimens such as membrane viruses in a near-native, frozen-hydrated state. These tomograms reveal structures of interest in three dimensions, albeit at low resolution. Computational averaging of sub-volumes, or sub-tomograms, is necessary to obtain higher resolution detail of repeating structural motifs, such as viral glycoprotein spikes. A detailed computational approach for aligning and averaging sub-tomograms using the Jsubtomo software package is outlined. This approach enables visualization of the structure of viral glycoprotein spikes to a resolution in the range of 20-40 Å and study of the study of higher order spike-to-spike interactions on the virion membrane. Typical results are presented for Bunyamwera virus, an enveloped virus from the family Bunyaviridae. This family is a structurally diverse group of pathogens posing a threat to human and animal health. |
| first_indexed | 2024-03-07T04:49:09Z |
| format | Journal article |
| id | oxford-uuid:d45dc0d1-aa8c-46a4-965f-d36d3b923ce4 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T04:49:09Z |
| publishDate | 2014 |
| publisher | Journal of Visualized Experiments |
| record_format | dspace |
| spelling | oxford-uuid:d45dc0d1-aa8c-46a4-965f-d36d3b923ce42022-03-27T08:17:54ZAveraging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d45dc0d1-aa8c-46a4-965f-d36d3b923ce4EnglishSymplectic Elements at OxfordJournal of Visualized Experiments2014Huiskonen, JParsy, MLi, SBitto, DRenner, MBowden, TEnveloped viruses utilize membrane glycoproteins on their surface to mediate entry into host cells. Three-dimensional structural analysis of these glycoprotein 'spikes' is often technically challenging but important for understanding viral pathogenesis and in drug design. Here, a protocol is presented for viral spike structure determination through computational averaging of electron cryo-tomography data. Electron cryo-tomography is a technique in electron microscopy used to derive three-dimensional tomographic volume reconstructions, or tomograms, of pleomorphic biological specimens such as membrane viruses in a near-native, frozen-hydrated state. These tomograms reveal structures of interest in three dimensions, albeit at low resolution. Computational averaging of sub-volumes, or sub-tomograms, is necessary to obtain higher resolution detail of repeating structural motifs, such as viral glycoprotein spikes. A detailed computational approach for aligning and averaging sub-tomograms using the Jsubtomo software package is outlined. This approach enables visualization of the structure of viral glycoprotein spikes to a resolution in the range of 20-40 Å and study of the study of higher order spike-to-spike interactions on the virion membrane. Typical results are presented for Bunyamwera virus, an enveloped virus from the family Bunyaviridae. This family is a structurally diverse group of pathogens posing a threat to human and animal health. |
| spellingShingle | Huiskonen, J Parsy, M Li, S Bitto, D Renner, M Bowden, T Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title | Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title_full | Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title_fullStr | Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title_full_unstemmed | Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title_short | Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo. |
| title_sort | averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using jsubtomo |
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