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3D Reconstruction of Mannosidase II from Single Particle Distributions: Noise Reduction Approaches for Higher Resolution

Published online by Cambridge University Press:  02 July 2020

F.P. Ottensmeyer ,
A.B. Fernandes ,
M. Timmer ,
J. Kroft ,
K. Varga  and
K.W. Moremen
F.P. Ottensmeyer
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G2M9, Canada
A.B. Fernandes
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G2M9, Canada
M. Timmer
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G2M9, Canada
J. Kroft
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G2M9, Canada
K. Varga
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G2M9, Canada
K.W. Moremen
Affiliation:
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA30602-7229, U.S.A.
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Extract

3D electron microscopy of macromolecules is now moving beyond the very successful tomographic approach of using images of a tilt-series of 2D crystals at known angles to single randomly oriented particles with less and less internal symmetry. The success of 3D reconstruction of symmetrical objects was predicated primarily on the redundancy of information due to lateral symmetry in 2D crystals, or to internal rotational symmetry in the icosahedral, helical or cylindrically symmetric particles. Such a spatially predictable redundancy permitted both a reduction in irradiation dose, to reduce radiationinduced structural alterations of the molecules, and an averaging of image noise due to the low number of electrons used and from the specimen support.

For 3D reconstruction from single particles the noise level in the images is crucial for the accurate determination of the relative orientation of the particles. In principle the central line theorem was recognized to be as applicable to asymmetric particles as it was to symmetrical viruses.

Type
Chambers and Channels: Functional Connections in Multiprotein Complexes Studied by Single Chambers and Channels
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 970 - 971
Copyright
Copyright © Microscopy Society of America

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