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The effect of protein-precipitant interfaces and applied shear on the nucleation and growth of lysozyme crystals

journal contribution
posted on 15.07.2014, 08:10 authored by Nuno Reis, D.Y. Chirgadze, T.L. Blundell, Malcolm R. Mackley
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.

History

School

  • Aeronautical, Automotive, Chemical and Materials Engineering

Department

  • Chemical Engineering

Published in

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY

Volume

65

Pages

1127 - 1139

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.

Publisher

© International Union of Crystallography

Version

AM (Accepted Manuscript)

Publication date

2009

Notes

This paper is closed access.

ISSN

0907-4449

eISSN

1399-0047

Language

en