Chapter 3.1 Preparing recombinant proteins for X-ray crystallography

It is possible to use recombinant DNA techniques to produce most proteins in quantities sufficient for crystallography. This chapter discusses issues (and problems) relevant to choosing methods appropriate for preparing recombinant proteins for X-ray crystallography. The aim is to help readers understand both the extant problems and the available solutions, so that, armed with a general understanding of the issues, they can more easily confront a variety of specific projects. Specific topics covered include protein engineering an expression construct, expression systems, protein purification and the characterization of the purified product.

Keywords: E. coli expression systems; affinity chromatography; baculovirus expression systems; chromatography; expression systems; insect cell–virus expression systems; mammalian-cell expression systems; protein expression; protein folding; protein purification; protein refolding; storage of proteins; tags; yeasts as expression systems