CRYSTAL-GROWTH AND CRYSTAL IMPROVEMENT STRATEGIES

The diffraction resolution of crystals of the guanine nucleotide-exchange factor complex, EF-Tu-Ts, has been extended from 5.0 to 2.5 angstrom by lowering the solvent content in the, crystals as well as the temperature of data collection. The common form of EF-Tu-Ts crystal belongs to space group P2(1)2(1)2(1) with a = 81.1, b 109.9, c = 207.5 angstrom and has a solvent content of 61 The crystals diffract to a resolution of 5.0 angstrom at 293 K and 4.0 angstrom at 273 K. When cryoprotective agents are slowly diffused into the crystals, the cell constants shrink to a = 74.4, b = 109.9, c = 198.7 angstrom and the solvent content falls to 55%. After the cryoprotective agent has been added, the crystals diffract to 2.7 angstrom resolution at 293 or 273K and 2.5 angstrom at 250 K. X-ray diffraction data, collected before and after the transformation of individual EF-Tu-Ts crystals, demonstrate that a large percentage of the improvement in diffraction resolution is due solely to the addition of cryoprotective agents. The transfer procedures for the successful introduction of cryoprotective agents into EF-Tu-Ts crystals as well as the general applicability to other crystal systems will be discussed.