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Please use this identifier to cite or link to this item: https://dspace.lboro.ac.uk/2134/15206

Title: The effect of protein-precipitant interfaces and applied shear on the nucleation and growth of lysozyme crystals
Authors: Reis, Nuno M.
Chirgadze, D.Y.
Blundell, T.L.
Mackley, Malcolm R.
Keywords: Lysozyme crystallization
Concentration gradients
Issue Date: 2009
Publisher: © International Union of Crystallography
Citation: REIS, N.M. ... et al, 2009. The effect of protein-precipitant interfaces and applied shear on the nucleation and growth of lysozyme crystals. Acta Crystallographica Section D - Biological Crystallography, 65, pp.1127-1139.
Abstract: This paper is concerned with the effect of protein–precipitant interfaces and externally applied shear on the nucleation and growth kinetics of hen egg-white lysozyme crystals. The early stages of microbatch crystallization of lysozyme were explored using both optical and confocal fluorescence microscopy imaging. Initially, an antisolvent (precipitant) was added to a protein drop and the optical development of the protein– precipitant interface was followed with time. In the presence of the water-soluble polymer poly(ethylene glycol) (PEG) a sharp interface was observed to form immediately within the drop, giving an initial clear separation between the lighter protein solution and the heavier precipitant. This interface subsequently became unstable and quickly developed within a few seconds into several unstable ‘fingers’ that represented regions of high concentration-gradient interfaces. Confocal microscopy demonstrated that the subsequent nucleation of protein crystals occurred preferentially in the region of these interfaces. Additional experiments using an optical shearing system demonstrated that oscillatory shear significantly decreased nucleation rates whilst extending the growth period of the lysozyme crystals. The experimental observations relating to both nucleation and growth have relevance in developing efficient and reliable protocols for general crystallization procedures and the controlled crystallization of single large high-quality protein crystals for use in X-ray crystallography.
Description: This paper is closed access.
Version: Accepted for publication
DOI: 10.1107/S0907444909031527
URI: https://dspace.lboro.ac.uk/2134/15206
Publisher Link: http://dx.doi.org/10.1107/S0907444909031527
ISSN: 0907-4449
Appears in Collections:Closed Access (Chemical Engineering)

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