title: Radiation damage to protein specimens from electron beam imaging and diffraction: a mini-review of anti-damage approaches, with special reference to synchrotron X-ray crystallography.
abstract: Recent research progress using X-ray cryo-crystallography with the photon beams from third-generation synchrotron sources has resulted in recognition that this intense radiation commonly damages protein samples even when they are held at 100 K. Other structural biologists examining thin protein crystals or single particle specimens encounter similar radiation damage problems during electron diffraction and imaging, but have developed some effective countermeasures. The aim of this concise review is to examine whether analogous approaches can be utilized to alleviate the X-ray radiation damage problem in synchrotron macromolecular crystallography. The critical discussion of this question is preceded by presentation of background material on modern technical procedures with electron beam instruments using 300-400 kV accelerating voltage, low-dose exposures for data recording, and protection of protein specimens by cryogenic cooling; these practical approaches to dealing with electron radiation damage currently permit best resolution levels of 6 A (0.6 nm) for single particle specimens, and of 1.9 A for two-dimensional membrane protein crystals. Final determination of the potential effectiveness and practical value of using such new or unconventional ideas will necessitate showing, by experimental testing, that these produce significantly improved protection of three-dimensional protein crystals during synchrotron X-ray diffraction.
authors.affiliation: Department of Cell Biology and Molecular Medicine, UMDNJ - New Jersey Medical School, Newark, NJ 07101-1709, USA. massover@andromeda.rutgers.edu
authors.forename: William H
authors.initials: WH
authors.lastname: Massover
mesh_terms: D003198:Computer Simulation; D018360:Crystallography, X-Ray / Q000379:methods*; D004307:Dose-Response Relationship, Radiation; D004583:Electrons; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation / Q000528:radiation effects; D011489:Protein Denaturation / Q000528:radiation effects; D011506:Proteins / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D011829:Radiation Dosage; D017356:Synchrotrons; D014965:X-Rays
chemical_list: D011506:Proteins