Abstract
The knowledge of accurate molecular structures obtained by X-ray protein crystallography is now inevitable for rational drug design and for understanding the molecular basis underlying genetic disorders found in patients. However, preparing protein crystals suitable for structural analysis is currently the bottleneck in structure determination by this method. The intent of this chapter is to present current methods of preparing protein crystals for structural studies for a wide range of biologists who have access to macromolecules but do not know how to handle them for crystallization. The chapter includes the pretreatment of a protein prior to the crystallization experiment, initial screens, and optimization of the crystallization conditions for further X-ray study. Finally, handling considerations that are important for a protein intended for crystallization experiments are discussed.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Drenth, J. (1994) Principles of Protein X-Ray Crystallography. (Cantor, C.R., ed.), Springer-Verlag, New York.
Blundell, T. L. and Johnson, L. N. (eds.) (1976) Protein Crystallography. Academic, Inc., San Diego.
Blow, D. (ed.) (2002) Outline of Crystallography for Biologists. Oxford University Press, New York.
Ducruix, A. and Giege, R. (eds.) (1999) Crystallization of Nucleic Acids and Proteins: A Practical Approach. Oxford University Press, New York.
McPherson, A. (ed.) (1999) Crystallization of Biological Macromolecules. Cold Spring Harbor Laboratory, New York.
Terese, M. B. (ed.) (1999) Protein Crystallization: Techniques, Strategies, and Tips. International University Line, La Jolla.
McRee, D. E. (ed.) (1999) Practical Protein Crystallography. Academic, San Diego.
Jancarik, J. and Kim, S. H. (1991) Sparse matrix sampling: a screening method for crystallization of protein. J. Appl. Cryst. 24, 409–411.
Vassylyev, D. G., Takeda, S., Wakatsuki, S., Maeda, K., and Maeda, Y. (1998) Crystal structure of troponin C in complex with troponin I fragment at 2.3-A resolution. Proc. Natl. Acad. Sci. USA 95, 4847–4852.
Takeda, S., Yamashita, A., Maeda, K., and Maeda, Y. (2003) Structure of the core domain of human cardiac troponin in the Ca(2+)-saturated form. Nature 424, 35–41.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Takeda, S., Kobayashi, T., Taniguchi, H., Hayashi, H., and Maeda, Y. (1997) Structural and functional domains of the troponin complex revealed by limited digestion. Eur. J. Biochem. 246, 611–617.
Pace, C. N., Vajdos, F., Fee, L., Grimsley, G., and Gray, T. (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci. 4, 2411–2423.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Takeda, S. (2006). Preparation of Protein Crystals for X-Ray Structural Study. In: Wang, Q.K. (eds) Cardiovascular Disease. Methods in Molecular Medicine, vol 129. Humana Press. https://doi.org/10.1385/1-59745-213-0:291
Download citation
DOI: https://doi.org/10.1385/1-59745-213-0:291
Publisher Name: Humana Press
Print ISBN: 978-1-58829-892-8
Online ISBN: 978-1-59745-213-7
eBook Packages: Springer Protocols