Crystallizing a Protein
Students new to the protein X-ray crystallography laboratory may understandably be confused when colleagues discuss Fouriers and Pattersons or molecular replacement and molecular dynamics refinement. However, they understand immediately that the first requirement for protein structure determination is to grow suitable crystals. Without crystals there can be no X-ray structure determination of a protein! In this chapter we discuss the principles of protein crystal growth and as an exercise give the recipe for crystallizing the enzyme lysozyme. We shall also generate an X-ray diffraction picture of a lysozyme crystal. This will provide an introduction to X-ray diffraction. The chapter concludes with a discussion of the problems encountered.
KeywordsProtein Solution Mother Liquor Reciprocal Lattice Protein Crystal Crystallization Experiment
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- 1.Protein crystallization is extensively discussed in: Ducruix, A. and Giegé, R. (1992). For the crystallization of membrane proteins, see Michel, H. (1990) and Sowadski (1994).Google Scholar
- 2.Hampton Research, 27632 El Lazo Road, Suite 100, Laguna Niguel, CA 92677 USA. Tel.: 1–714-425–1321; Fax: 1–714-425–1611, http://www.hamptonresearch.com; e-mail: firstname.lastname@example.org
- 3.Paul P. Ewald, 1888–1985, professor of physics at several universities in Europe and the United States, was the first to apply the reciprocal lattice and the sphere named after him to the interpretation of an X-ray diffraction pattern.Google Scholar