Bulk preparation and crystallization of the Escherichia coli elongation factor Tu-Ts complex

Abstract

A simple procedure for the preparation of 10-500 mg of the Escherichia coli elongation Tu-Ts complex is described. The protocol is based on the separate purification and quantitation of EF-Tu-GDP and EF-Ts, followed by mixing of equimolar amounts of each protein and removal of the displaced GDP by dialysis. Single crystals grown from the final product have been analyzed by X-ray diffraction techniques. The procedure is also applicable to the bulk preparation and crystallization of the trypsin-modified Tu-Ts complex. Quantitation of the elongation factors by three methods is presented.