Plastic-embedded protein crystals

R. Ravelli, U. Haselmann-Weiss, John McGeehan, A. McCarthy, J. Marquez, C. Antony, A. Frangakis, G. Stranzl

Research output: Contribution to journalArticlepeer-review

Abstract

Rapid vitrification followed by the replacement of the vitrified water by a solvent (freeze substitution) and then resin is a widely used procedure for preparing biological samples for electron microscopy. The resulting plastic-embedded samples permit convenient room-temperature sectioning (microtomy) and can yield well preserved cellular structures. Here this procedure has been applied to crystalline protein samples, and it is shown that it is possible to freeze-substitute vitrified crystals while preserving some of their original diffraction properties. The plastic-embedded crystals were used to collect a series of complete room-temperature data sets at a powerful macromolecular crystallography synchrotron beamline. Whereas one normally observes specific damage to disulfide bonds upon X-ray radiation, no such damage was seen for the plastic-embedded sample. The X-ray diffraction data allowed an initial atomic analysis to be made of the effects of freeze-substitution and plastic embedding on biological samples.
Original languageEnglish
Pages (from-to)128-132
Number of pages5
JournalJournal of Synchrotron Radiation
Volume14
Issue number1
DOIs
Publication statusPublished - 2007

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