The crystallization step is considered to be a major bottleneck in the process of structure determination by x-ray crystallography. Successful crystallization requires both the formation of nuclei that are capable of supporting crystal growth, and a subsequent crystal growth. Nucleation can occur spontaneously in a supersaturated solution. However, in the commonly used sparse matrix crys tallization screens, protein and precipitant concentrations are not extensively sampled, so that suitable supersaturation conditions for nucleation are often missed. This chapter describes a simple method for enhancing nucleation and subsequent crystallization by the addition of heterogeneous nucleating agents.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McPherson, A. (2004) Introduction to protein crystallization. Methods 34, 254–265.
McPherson, A. (1999) Crystallization of Biological Macromolecules, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Asherie, N. (2004) Protein crystallization and phase diagrams. Methods 34, 266–272.
Rong, L., Komatsu, H., Yoshizaki, I., Kadowaki, A., and Yoda, S. (2004) Protein crystallization by using porous glass substrate. J. Synchrotron Radiat. 11, 27–29.
Yoshizaki, I., Sato, T., Igarashi, N., Natsuisaka, M., Tanaka, N., Komatsu, H., and Yoda, S. (2001) Systematic analysis of supersaturation and lysozyme crystal quality. Acta Crystallogr. D Biol. Crystallogr. 57, 1621–1629.
McPherson, A., and Schlichta, P. (1988) Heterogeneous and epitaxial nucleation of protein crystals on mineral surfaces. Science 239, 385–387.
Bergfors, T. (2003) Seeds to crystals. J. Struct. Biol. 142, 66–76.
McPherson, A., and Shlichta, P. J. (1987) Facilitation of the growth of protein crys tals by heterogeneous/epitaxial nucleation. J. Cryst. Growth 85, 206–214.
Edwards, A. M., Darst, S. A., Hemming, S. A., Li, Y., and Kornberg, R. D. (1994) Epitaxial growth of protein crystals on lipid layers. Nature Struct. Biol. 1, 195–197.
Punzi, J. S., Luft, J., and Cody, V. (1991) Protein crystal growth in the presence of poly(vinylidene diflouride) membrane. J. Appl. Cryst. 24, 406–408.
Chayen, N. E., Shaw Stewart, P. D., Maeder, D. L., and Blow, D. M. (1990) An automated system for micro-batch protein crystallization and screeing. J. Appl. Crytallogr. 23, 297–302.
Pechkova, E., and Nicolini, C. (2002) Protein nucleation and crystallization by homologous protein thin film template. J. Cell Biochem. 85, 243–251.
Chayen, N. E., Saridakis, E., El-Bahar, R., and Nemirovsky, Y. (2001) Porous silicon: an effective nucleation-inducing material for protein crystallization. J. Mol. Biol. 312, 591–595.
Hench, L. L. (1998) Biomaterials: a forecast for the future. Biomaterials 19, 1419–1423.
Fermani, S., Falini, G., Minnucci, M., and Ripamonti, A. (2001) Protein crystallization on polymeric film surfaces J. Cryst. Growth 224, 327–334.
Leung, C. J., Nall, B. T., and Brayer, G. D. (1989) Crystallization of yeast iso-2-cytochrome c using a novel hair seeding technique. J. Mol. Biol. 206, 783–785.
Stura, E. A., and Wilson, I. A. (1992), Seeding techniques in (Ducruix, A., and Giegé, R., eds.), Crystallization of Nucleic Acids and Proteins: A Practical Approach, pp. 99–126, Oxford University Press, Oxford, UK.
Jancarik, J., and Kim, S.-H. (1991) Sparse matrix sampling: a screening method for crystallization of proteins. J. Appl. Cryst. 24, 409–411.
Thakur, A. S., Robin, G., Guncar, G., Saunders, N. F., Newman, J., Martin, J. L., and Kobe, B. Improved success of protein crystallization sparse matrix screening with heterogeneous nucleating agents. PLoS ONE.
Chernov, A. A. (2003) Protein crystals and their growth. J. Struct. Biol. 142, 3–21.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Thakur, A.S., Newman, J., Martin, J.L., Kobe, B. (2008). Increasing Protein Crystallization Screening Success with Heterogeneous Nucleating Agents. In: Kobe, B., Guss, M., Huber, T. (eds) Structural Proteomics. Methods in Molecular Biology™, vol 426. Humana Press. https://doi.org/10.1007/978-1-60327-058-8_26
Download citation
DOI: https://doi.org/10.1007/978-1-60327-058-8_26
Publisher Name: Humana Press
Print ISBN: 978-1-58829-809-6
Online ISBN: 978-1-60327-058-8
eBook Packages: Springer Protocols