International
Tables for Crystallography Volume C Mathematical, physical and chemical tables Edited by E. Prince © International Union of Crystallography 2006 |
International Tables for Crystallography (2006). Vol. C. ch. 3.4, p. 166
|
Crystals of biological macromolecules are very susceptible to radiation damage, and this can severely limit the amount and quality of diffraction data that can be collected per crystal. There have been relatively few systematic studies of this phenomenon (Young, Dewan, Nave & Tilton, 1993; Gonzalez & Nave, 1994
; Nave, 1995
), but one of the first effects of radiation damage is the deterioration of the high-resolution regions of the pattern, followed by increasing loss of crystallinity. Improvement of crystal lifetime in X-ray beams has been obtained by the addition of free-radical scavengers (Zaloga & Sarma, 1974
) and the replacement of the mother liquor with solutions containing 10–20% polyethylene glycol 4000 or 20000 (Cascio, Williams & McPherson, 1984
). The use of synchrotron radiation has also led to improved data-per-crystal ratios (Lindley, 1988
). The high intensity allows fast collection of data, and the high collimation permits different sections of the same crystal to be used for data collection. This is particularly useful for prismatic crystals, which can be mounted along their largest morphological axis. An alternative method of surmounting this problem, however, is to freeze the protein crystal. As the temperature is decreased, the rate of diffusion of free radicals is reduced, with a corresponding reduction in radiation damage. Appreciable reduction in diffusion rate is achieved even at 250 K, and at 100 K diffusion essentially ceases. Cryogenic measurements not only minimize radiation damage but often lead to improved resolution owing to decrease in thermal motion in the crystal. Increasing the crystal lifetime may be particularly important with respect to multiwavelength anomalous-dispersion measurements in order to derive phase information. Since crystals of biological macromolecules contain substantial amounts of solvent, typically between 35 and 80% by volume, the technical problem is to force the solvent to cool in an amorphous glass-like state, rather than as crystalline ice. The latter normally degrades the crystallinity by expansion and gives rise to powder rings, which complicate data measurement.
References





