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Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging

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

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41,150 Views

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07:29 min

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December 01, 2011

DOI:

10.3791/2770-v

DOI:
10.3791/2770-v

•

07:29 min
December 01, 2011

•

1 Views
, , , , , , , , , , , , , , , , , , ,
1Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute,National Institutes of Health, 2The Medical Research Council Mitochondrial Biology Unit,University of Cambridge , 3National Library of Medicine,National Institutes of Health, 4Massachusetts Institute of Technology, 5William Fremd High School, 6University of Virginia , 7Duke University , 8Yale University, 9University of Notre Dame , 10Washington University in St. Louis , 11Bioinformatics and Computational Biosciences Branch, National Institute of Allergy and Infectious Diseases,National Institutes of Health, 12Thomas Jefferson High School for Science and Technology

Summary

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The protocol describes a high-throughput approach to determining structures of membrane proteins using cryo-electron tomography and 3D image processing. It covers the details of specimen preparation, data collection, data processing and interpretation, and concludes with the production of a representative target for the approach, the HIV-1 Envelope glycoprotein. These computational procedures are designed in a way that enables researchers and students to work remotely and contribute to data processing and structural analysis.

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Meyerson, J. R., White, T. A., Bliss, D., Moran, A., Bartesaghi, A., Borgnia, M. J., de la Cruz, M. J. V., Schauder, D., Hartnell, L. M., Nandwani, R., Dawood, M., Kim, B., Kim, J. H., Sununu, J., Yang, L., Bhatia, S., Subramaniam, C., Hurt, D. E., Gaudreault, L., Subramaniam, S. Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging. J. Vis. Exp. (58), e2770, doi:10.3791/2770 (2011).

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