Phasing macromolecular structures with UV-induced structural changes.
Experimental phasing of macromolecular crystal structures relies on the accurate measurement of two or more sets of reflections from isomorphous crystals, where the scattering power of a few atoms is different for each set. Recently, it was demonstrated that X-ray-induced intensity differences can also contain phasing information, exploiting specific structural changes characteristic of X-ray damage. This method (radiation damage-induced phasing; RIP) has the advantage that it can be performed on a single crystal of the native macromolecule. However, a drawback is that X-rays introduce many small changes to both solvent and macromolecule. In this study, ultraviolet (UV) radiation has been used to induce specific changes in the macromolecule alone, leading to a larger contrast between radiation-susceptible and nonsusceptible sites. Unlike X-ray RIP, UV RIP does not require the use of a synchrotron. The method has been demonstrated for a series of macromolecules.
Nanao
Max H
MH
European Molecular Biology Laboratory (EMBL), Grenoble Outstation, 6 rue Jules Horowitz, B.P. 181, 38042 Grenoble Cedex 9, France.
Ravelli
Raimond B G
RB
D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004220:Disulfides; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry*; D008958:Models, Molecular; D015233:Models, Statistical; D008968:Molecular Conformation; D015394:Molecular Structure; D011485:Protein Binding; D011487:Protein Conformation; D011506:Proteins / Q000528:radiation effects*; D013053:Spectrophotometry; D017356:Synchrotrons; D014466:Ultraviolet Rays
D004220:Disulfides; D046911:Macromolecular Substances; D011506:Proteins