X-ray diffraction studies of protein crystal disorder

Protein crystals contain many kinds of disorder, but only a small fraction of these are likely to be important in limiting the diffraction properties of interest to crystallographers. X-ray topography, high-angular-resolution reciprocal space measurements, and standard crystallographic data collection have been used to probe three factors that may produce diffraction-limiting disorder: (1) solution variations during crystal growth, (2) macromolecular impurities, and (3) post-growth crystal treatments. Variations in solution conditions that occur in widely used growth methods may lead to variations in equilibrium protein conformation and crystal packing as a crystal grows, and these may introduce appreciable disorder for sensitive proteins. Tetragonal lysozyme crystals subjected to abrupt changes in temperature, pH, or salt concentration during growth show increased disorder, consistent with this mechanism. Macromolecular impurities can have profound effects on protein crystal quality. A combination of diffraction measurements provides insight into the mechanisms by which particular impurities create disorder, and this insight leads to a simple approach for reducing this disorder. Substantial degradation of diffraction properties due to conformation and lattice constant changes can occur during post-growth crystal treatments such as heavy-atom compound and drug binding. Measurements of the time evolution of crystal disorder during controlled crystal dehydration — a simple model for such treatments — suggest that structural metastability conferred by the constraints of the crystal lattice plays an important role in determining the extent to which the diffraction properties degrade. ( 1999 Elsevier Science B.V. All rights reserved. PACS: 61.72; 81.10; 87.15