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Three-Dimensional Structure Of C3D From Low Temperature Scanning Transmission Electron Microscopy and X-Ray Crystallography

Published online by Cambridge University Press:  02 July 2020

D.J. Martin  and
F.P. Ottensmeyer
D.J. Martin
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada, M5G 2M9
F.P. Ottensmeyer
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada, M5G 2M9
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Macromolecular structure can be solved by x-ray crystallography to atomic resolution provided that the molecule can be crystallized, that the crystals diffract x-rays to high resolution, and that the phases of the diffracted x-rays can be determined. Though the resolution of single particle imaging by electron microscopy is lower than that of x-ray diffraction by crystals, electron microscopy can directly image a large molecular weight range of macromolecules in a non-crystalline environment, and provide the basis for the three-dimensional reconstruction of these structures. To investigate combining structural information from x-ray crystallography and electron microscopy for unknown structures, we have imaged a small protein of known structure (1), the 35 kDa human complement protein fragment C3d, in a scanning transmission electron microscope (STEM). The intention is to eventually combine the knowledge of electron densities and molecular boundaries from electron microscopy to assist in phase determination in x-ray crystallography.

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. 294 - 295
Copyright
Copyright © Microscopy Society of America

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References

REFERENCES

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6. This work was supported by funds from NSERC of Canada and MRC of Canada. The donation of C3d by Dr. James Rini of the University of Toronto is greatly appreciated.Google Scholar