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Examining Protein Crystallization Using Scanning Electron Microscopy

Published online by Cambridge University Press:  30 November 2012

Kathryn Gomery ,
Elaine C. Humphrey  and
Rodney Herring
Kathryn Gomery*
Affiliation:
University of Victoria, Department of Biochemistry and Microbiology, P.O. Box 3055 STN CSC, Victoria, BC V8W 3P6, Canada
Elaine C. Humphrey
Affiliation:
University of Victoria, Department of Mechanical Engineering, P.O. Box 3055 STN CSC, Victoria, BC V8W 3P6, Canada
Rodney Herring
Affiliation:
University of Victoria, Department of Mechanical Engineering, P.O. Box 3055 STN CSC, Victoria, BC V8W 3P6, Canada
*
*Corresponding author. E-mail: [email protected]
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Abstract

Elucidation of protein structure using X-ray crystallography relies on the quality of the crystal. Crystals suffer from many different types of disorder, some of which occur during crystal nucleation and early crystal growth. To date, there are few studies surrounding the quality and nucleation of protein crystals partly due to difficulties surrounding viewing biological samples at high resolution. Recent research has led our current understanding of nucleation to be a two-step mechanism involving the formation of nuclei from dense liquid clusters; it is still unclear whether nuclei first start as amorphous aggregate or as crystalline lattices. Our research examines this mechanism through the use of electron microscopy. Using scanning electron microscopy imaging of the protein crystal growth process, a stacking, spiraling manner of growth is observed. The tops of the pyramid-like tetragonal protein crystal structures measure ~0.2 μm across and contain ~125,000 lysozyme units. This noncrystalline area experiences strain due to growth of the protein crystal. Our work shows that it is possible to view detailed early stage protein crystal growth using a wet scanning electron microscopy technique, thereby overcoming the problem of viewing liquids in a vacuum.

Keywords

protein crystallographylysozymeSEMnucleation
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 19 , Issue 1 , February 2013 , pp. 145 - 149
Copyright
Copyright © Microscopy Society of America 2013

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