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Vive La Différence! Mapping Macromolecular Complexes by Generalized Difference Imaging

Published online by Cambridge University Press:  02 July 2020

Alasdair C. Steven ,
James F. Conway ,
Naiqian Cheng ,
Norman R. Watts  and
Paul T. Wingfield
Alasdair C. Steven
Affiliation:
Laboratory of Structural Biology and Protein Expression Laboratory, NIAMS-NIH, Bethesda, MD20892;
James F. Conway
Affiliation:
Laboratory of Structural Biology and Protein Expression Laboratory, NIAMS-NIH, Bethesda, MD20892; Protein Expression Laboratory, NIAMS-NIH, Bethesda, MD20892;
Naiqian Cheng
Affiliation:
Laboratory of Structural Biology and Protein Expression Laboratory, NIAMS-NIH, Bethesda, MD20892;
Norman R. Watts
Affiliation:
Laboratory of Structural Biology and Protein Expression Laboratory, NIAMS-NIH, Bethesda, MD20892; Institut de Biologie Structural, 41 rue Jules Horowitz, 38027 Grenoble Cedex 1, France
Paul T. Wingfield
Affiliation:
Institut de Biologie Structural, 41 rue Jules Horowitz, 38027 Grenoble Cedex 1, France
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Extract

Overview. Three-dimensional cryo-electron microscopy holds great potential for the investigation of macromolecular complexes, including viruses, the cytoskeleton, and the nanomachines that carry out many vital cellular functions. Such complexes often contain protein subunits of different kinds, with cumulative molecular weights running well into the megadalton range. To rationalize their functional assignments in structural terms, the first order of business is to map the locations of the component subunits of a given complex. A more detailed objective then becomes to understand the organization of their domains and the overall layout of the polypeptide chains. With current methodology, resolutions below 20Å are often achievable in cryo-EM, extending below 10 Å in favorable cases. Although resolutions of this order yield valuable morphological information, they do not usually suffice to identify individual subunits within a complex. Generalized difference imaging offers a solution to this problem of molecular mapping.

Type
Electron Cryomicroscopy of Macromolecules
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 252 - 253
Copyright
Copyright © Microscopy Society of America

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