Crystallization of soluble proteins in vapor diffusion for x-ray crystallography.
The preparation of protein single crystals represents one of the major obstacles in obtaining the detailed 3D structure of a biological macromolecule. The complete automation of the crystallization procedures requires large investments in terms of money and labor, which are available only to large dedicated infrastructures and is mostly suited for genomic-scale projects. On the other hand, many research projects from departmental laboratories are devoted to the study of few specific proteins. Here, we try to provide a series of protocols for the crystallization of soluble proteins, especially the difficult ones, tailored for small-scale research groups. An estimate of the time needed to complete each of the steps described can be found at the end of each section.
Benvenuti
Manuela
M
Dipartimento di Chimica, Università di Siena, Via Aldo Moro 2, Siena 53100, Italy.
Mangani
Stefano
S
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004058:Diffusion; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012995:Solubility
D011506:Proteins