Name: Determinação estrutural de hemaglutinina de resolução subnanométrica a partir de tomografia crioeletrônica de vírus influenza (Vídeo) | JoVE Journal
Uploaded: 2025-11-07T12:50:00.000+00:00
Duration: 8 min 19 s
Description: Aprenda a usar tomografia crioeletônica para visualizar a hemaglutinina do vírus influenza com resolução subnanométrica. Este protocolo permite estudar mudanças conformacionais durante a ligação ao receptor e analisar a organização viral in situ.

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</ol>

<table><tbody><tr><td>AMD Ryzen Threadripper PRO 5965WX</td><td>AMD</td><td>https://www.amd.com/en/support/downloads/drivers.html/processors/ryzen-threadripper-pro/ryzen-threadripper-pro-5000wx-series/amd-ryzen-threadripper-pro-5965wx.html</td><td></td></tr><tr><td>AreTomo 1.3.4</td><td>UC San Francisco</td><td>https://drive.google.com/drive/folders/1Z7pKVEdgMoNaUmd_cOFhlt-QCcfcwF3_</td><td></td></tr><tr><td>EMAN2 2.99.52</td><td>Faculdade de Medicina Baylor</td><td>https://blake.bcm.edu/emanwiki/EMAN2</td><td></td></tr><tr><td>IMOD 4.12.27</td><td>Universidade do Colorado em Boulder</td><td>https://bio3d.colorado.edu/imod/</td><td></td></tr><tr><td>Roteiros de análise de influenza</td><td>Faculdade de Medicina UMass Chan</td><td>https://github.com/jqyhuang/influenza-analysis</td><td></td></tr><tr><td>IsoNet 0.3</td><td>UCLA</td><td>https://github.com/IsoNet-cryoET/IsoNet</td><td></td></tr><tr><td>M 2.0.0</td><td>Genentech</td><td>https://warpem.github.io/warp/home/m/</td><td></td></tr><tr><td>NVIDIA A4000</td><td>NVIDIA&nbsp;</td><td>https://www.nvidia.com/en-us/products/workstations/rtx-a4000/</td><td></td></tr><tr><td>Open3D&nbsp;</td><td>Intel Labs</td><td>https://www.open3d.org/</td><td></td></tr><tr><td>PHENIX 1.21-5207</td><td>Laboratório Nacional Lawrence Berkeley</td><td>phenix-online.org</td><td></td></tr><tr><td>RELION 4.0</td><td>Laboratório de Biologia Molecular do MRC</td><td>https://relion.readthedocs.io/en/release-4.0/</td><td></td></tr><tr><td>Ubuntu 20.04</td><td>Ubuntu</td><td>https://releases.ubuntu.com/focal/</td><td></td></tr><tr><td>UCSF ChimeraX 1.6.1</td><td>UC San Francisco</td><td>https://www.cgl.ucsf.edu/chimerax/</td><td></td></tr><tr><td>Warp 2.0.0</td><td>Genentech</td><td>http://warpem.github.io/warp/</td><td></td></tr></tbody></table>

determinação estrutural de hemaglutinina de resolução subnanométrica a partir de tomografia crioeletrônica de vírus influenza

Subnanometer-Resolution Structural Determination of Hemagglutinin from Cryo-Electron Tomography of Influenza Viruses

Determinação estrutural de hemaglutinina de resolução subnanométrica a partir de tomografia crioeletrônica de vírus influenza

Cryo-electron tomography is a powerful tool to visualize heterogeneous samples, with one major application being structural characterization of ...

subnanometer-resolution-structural-determination-hemagglutinin-from

Research

JoVE Journal

Biochemistry

Bioquímica

O protocolo descreve uma abordagem de alto rendimento para determinação de estruturas de proteínas de membrana usando tomografia crio-eletrônica e processamento de imagens 3D. Abrange os detalhes da preparação da amostra, coleta de dados, processamento e interpretação dos dados, e conclui com a produção de um alvo representante para a abordagem, o HIV-1 glicoproteína do envelope. Estes procedimentos computacionais são projetados de uma forma que permite aos pesquisadores e estudantes para...

Determinação de estruturas moleculares de glicoproteínas do envelope HIV usando Cryo-Electron Tomografia e Média Sub-tomogram Automated

Nós descrevemos um método de microscopia correlativa que combina alta velocidade de microscopia de luz fluorescente de células vivas 3D de alta resolução tomografia crio-eletrônico. Nós demonstrar a capacidade do método dinâmico de imagem por correlativo, as pequenas partículas de HIV-1 que interagem com células HeLa hospedeiras.

Microscopia correlativo para Análise Estrutural 3D de interações dinâmicas

Nós apresentamos um protocolo de como coletar e eletrônica processo de crio-tomograms de toda mitocôndrias. A técnica fornece informações detalhadas sobre a estrutura, função e organização de grandes complexos de proteínas de membrana em membranas biológicas nativas.

Visualização da ATP sintase Dimers na mitocôndria por Electron Cryo-tomografia

A fixação de virions a uma superfície é um requisito para imagens de virion única por imagens de fluorescência de super resolução ou microscopia de força atômica (AFM). Aqui demonstramos um método de preparação de amostras para adesão controlada de virions a superfícies de vidro adequadas para uso em AFM e imagens de fluorescência de super resolução.

Preparação da amostra para microscopia da força atômica de virion único e imagem de fluorescência de super-resolução

Uma abordagem é apresentada para determinar estruturas de complexos de glicoproteínas de membrana viral usando uma combinação de criotomografia eletrônica e média de subtomograma com o pacote computacional Jsubtomo.

Calculando a média de Viral glicoproteína do envelope Spikes de Electron Cryotomography reconstruções com Jsubtomo

Este artigo apresenta um protocolo para processamento de imagens cryo-EM usando o software suite SPHIRE. O presente protocolo pode ser aplicado para quase todos os projetos de EM de partículas individuais que visam a resolução quase atômica.

Análise de uma única partícula de alta resolução a partir de imagens de crio-microscopia de elétrons usando SPHIRE

Apresentamos um protocolo para a preparação de lamella de espécimes biológicos congelados por micromaquingia de feixe de íons focado em crio-pia para estudos estruturais de alta resolução de macromoléculas in situ com tomografia crio-elétron. O protocolo apresentado fornece diretrizes para a preparação de lamellae de alta qualidade com alta reprodutibilidade para caracterização estrutural de macromoléculas dentro do Saccharomyces cerevisiae.

Preparação e micromaquinção Crio-FIB de Saccharomyces cerevisiae para Tomografia Crio-Elétron

Aqui os protocolos são descritos para preparar conjuntos de vírus adequados para análise líquido-EM e crio-EM em nanoescala usando microscopia eletrônica de transmissão.

Avançando a imagem de alta resolução de conjuntos de vírus em líquido e gelo

O presente protocolo descreve a aquisição remota de dados remotos crio-elétrons de alta resolução usando Tomo5 e processamento de dados subsequente e média de subtomogramas usando a emClarity. A apoferritina é usada como exemplo para ilustrar processos passo a passo detalhados para alcançar uma estrutura crio-ET com resolução de 2,86 Å.

Coleta remota de dados de tomografia crio-elétron e média de subtomograma

Este protocolo de vídeo ilustra como realizar a tomografia por microscopia eletrônica de transmissão de varredura de espécimes virológicos. Para obter os melhores resultados, as amostras são preparadas por congelamento de alta pressão e subsequente substituição por congelamento.

Tomografia por Microscopia Eletrônica de Transmissão de Varredura em Virologia: Imagem 3D de Amostras Congeladas de Alta Pressão e Substituídas por Congelamento

Aprenda a usar tomografia crioeletônica para visualizar a hemaglutinina do vírus influenza com resolução subnanométrica. Este protocolo permite estudar mudanças conformacionais durante a ligação ao receptor e analisar a organização viral in situ.

Determinação estrutural de hemaglutinina de resolução subnanométrica a partir de tomografia crioeletrônica de vírus influenza

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