Bulk preparation and crystallization of the Escherichia coli elongation factor Tu-Ts complex.
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.
D002850:Chromatography, Gel; D003460:Crystallization; D003956:Dialysis; D004926:Escherichia coli / Q000032:analysis*; D006153:Guanosine Diphosphate / Q000032:analysis; D010444:Peptide Elongation Factor Tu / Q000302:isolation & purification*; D010445:Peptide Elongation Factors / Q000302:isolation & purification*; D011485:Protein Binding; D014357:Trypsin; D014961:X-Ray Diffraction
D010445:Peptide Elongation Factors; C023512:elongation factor Ts; D006153:Guanosine Diphosphate; D014357:Trypsin; D010444:Peptide Elongation Factor Tu