scholarly journals Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging

10.3791/2770 ◽  
2011 ◽  
Author(s):  
Joel R. Meyerson ◽  
Tommi A. White ◽  
Donald Bliss ◽  
Amy Moran ◽  
Alberto Bartesaghi ◽  
...  
Keyword(s):  
Electron Tomography ◽  
Molecular Structures ◽  
Envelope Glycoproteins ◽  
Cryo Electron Tomography ◽  
Hiv Envelope

scholarly journals Fine details in complex environments: the power of cryo-electron tomography

2018 ◽  
Vol 46 (4) ◽  
pp. 807-816 ◽  
Author(s):  
Joshua Hutchings ◽  
Giulia Zanetti
Keyword(s):  
Structural Information ◽  
Electron Tomography ◽  
Molecular Structures ◽  
Complex Environments ◽  
Single Particle Reconstruction ◽  
Wide Range ◽  
Cryo Electron Tomography ◽  
Recent Developments

Cryo-electron tomography (CET) is uniquely suited to obtain structural information from a wide range of biological scales, integrating and bridging knowledge from molecules to cells. In particular, CET can be used to visualise molecular structures in their native environment. Depending on the experiment, a varying degree of resolutions can be achieved, with the first near-atomic molecular structures becoming recently available. The power of CET has increased significantly in the last 5 years, in parallel with improvements in cryo-EM hardware and software that have also benefited single-particle reconstruction techniques. In this review, we cover the typical CET pipeline, starting from sample preparation, to data collection and processing, and highlight in particular the recent developments that support structural biology in situ. We provide some examples that highlight the importance of structure determination of molecules embedded within their native environment, and propose future directions to improve CET performance and accessibility.

scholarly journals 3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography

2012 ◽  
Vol 180 (2) ◽  
pp. 318-326 ◽  
Author(s):  
Ke Wang ◽  
Korrinn Strunk ◽  
Gongpu Zhao ◽  
Jennifer L. Gray ◽  
Peijun Zhang
Keyword(s):  
Structure Determination ◽  
Mammalian Cells ◽  
Electron Tomography ◽  
3D Structure ◽  
Cryo Electron Tomography ◽  
3D Structure Determination

scholarly journals In situ structure determination of virus capsids imaged within cell nuclei by correlative light and cryo-electron tomography

2020 ◽  
Author(s):  
Swetha Vijayakrishnan ◽  
Marion McElwee ◽  
Colin Loney ◽  
Frazer Rixon ◽  
David Bhella
Keyword(s):  
Electron Microscopy ◽  
Structure Determination ◽  
Electron Tomography ◽  
3D Structure ◽  
Three Dimensional ◽  
Cell Nuclei ◽  
Nuclear Egress ◽  
Virus Capsids ◽  
Cryo Electron Tomography

AbstractCryo electron microscopy (cryo-EM), a key method for structure determination involves imaging purified material embedded in vitreous ice. Images are then computationally processed to obtain three-dimensional structures at atomic resolution. There is increasing interest in extending structural studies by cryo-EM into the cell, where biological structures and processes may be imaged in context. The limited penetrating power of electrons prevents imaging of thick specimens (>500 nm) however. Cryo-sectioning methods employed to overcome this are technically challenging, subject to artefacts or involve specialised equipment of limited availability. Here we describe the first structure of herpesvirus capsids determined by sub-tomogram averaging from nuclei of eukaryotic cells, achieved by cryo-electron tomography (cryo-ET) of re-vitrified cell sections prepared using the Tokuyasu method. Our reconstructions reveal that the capsid associated tegument complex is present on capsids prior to nuclear egress. We show that this approach to cryogenic imaging of cells is suited to both correlative light/electron microscopy and 3D structure determination.

scholarly journals Understanding the mammalian TRAP complex function(s)

Open Biology ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 190244
Author(s):  
Antonietta Russo
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Tomography ◽  
Complex Function ◽  
Conducting Channel ◽  
Unfolded Protein ◽  
Cryo Electron Microscopy ◽  
Cryo Electron Tomography ◽  
Protein Response ◽  
Microscopic Techniques

In eukaryotic cells, about one-third of the synthesized proteins are translocated into the endoplasmic reticulum; they are membrane or lumen resident proteins and proteins direct to the Golgi apparatus. The co-translational translocation takes place through the heterotrimeric protein-conducting channel Sec61 which is associated with the ribosome and many accessory components, such as the heterotetrameric translocon-associated protein (TRAP) complex. Recently, microscopic techniques, such as cryo-electron microscopy and cryo-electron tomography, have enabled the determination of the translocation machinery structure. However, at present, there is a lack of understanding regarding the roles of some of its components; indeed, the TRAP complex function during co-translational translocation needs to be established. In addition, TRAP may play a role during unfolded protein response, endoplasmic-reticulum-associated protein degradation and congenital disorder of glycosylation (ssr4 CDG). In this article, I describe the current understanding of the TRAP complex in the light of its possible function(s).

scholarly journals Structural Studies of Trimeric SIV and HIV-1 Envelope Glycoproteins Using Cryo-Electron Tomography

2010 ◽  
Vol 16 (S2) ◽  
pp. 882-883
Author(s):  
TA White ◽  
A Bartesaghi ◽  
MJ Borgnia ◽  
MJV de la Cruz ◽  
JLS Milne ◽  
...  
Keyword(s):  
Electron Tomography ◽  
Envelope Glycoproteins ◽  
Structural Studies ◽  
Cryo Electron Tomography ◽  
Hiv 1

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

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