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Protein Characterization by Low-Angle, Rotary-Replication Electron Microscopy

Published online by Cambridge University Press:  02 July 2020

S. Samuelsson
S. Samuelsson*
Affiliation:
Research Analytical Section, Procter & Gamble Pharmaceuticals, Inc., 8700 Mason-Montgomery Road, Mason, OH45040-9462
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Extract

There are several established strategies for visualizing proteins by transmission electron microscopy (TEM). These include negative staining, cryo-TEM, glycerol-spray/rotary-replication, mica-flake/freeze-fracture. Of these, low-angle, rotary-replication of proteins dried out of glycerol has proven to be a reliable and accurate method for studying surface topologies of macromolecular assemblies and particles. We have characterized many proteins, both chimeras and wild type; methods, caveats and the surface structure of a couple examples are described here.

The technique of using heavy metals to cast replicas of proteins and particles has been around for decades (metal replication of proteins was first described in 1956) and continues to provide an excellent method for evaluating protein topology. The success of this method is based on the physical properties of protein in contact with freshly cleaved mica, glycerol and atomic platinum. Mica is easily split and atomically flat making it an excellent substrate.

Type
Microscopy and Microanalysis in the Pharmaceutical Industry
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. 994 - 995
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Hall, C.E.. Proc. Natl. Acad. Sci. 42 (1956) 801.CrossRefGoogle Scholar
2.Fowler, W.E. and Aebi, U.. J. Ultrastructural Research 83 (1983) 319.CrossRefGoogle Scholar
3.Hanahan, D.. Science 277 (1997) 48.CrossRefGoogle Scholar
4.Heuser, J.E.. J. Mol. Biol. 169 (1983) 155.CrossRefGoogle Scholar