title: Using X-ray absorption spectra to monitor specific radiation damage to anomalously scattering atoms in macromolecular crystallography.
abstract: Radiation damage in macromolecular crystals is not suppressed even at 90 K. This is particularly true for covalent bonds involving an anomalous scatterer (such as bromine) at the 'peak wavelength'. It is shown that a series of absorption spectra recorded on a brominated RNA faithfully monitor the extent of cleavage. The continuous spectral changes during irradiation preserve an 'isosbestic point', each spectrum being a linear combination of 'zero' and 'infinite' dose spectra. This easily yields a good estimate of the partial occupancy of bromine at any intermediate dose. The considerable effect on the near-edge features in the spectra of the crystal orientation versus the beam polarization has also been examined and found to be in good agreement with a previous study. Any significant influence of the (C-Br bond/beam polarization) angle on the cleavage kinetics of bromine was also searched for, but was not detected. These results will be useful for standard SAD/MAD experiments and for the emerging 'radiation-damage-induced phasing' method exploiting both the anomalous signal of an anomalous scatterer and the 'isomorphous' signal resulting from its cleavage.
authors.affiliation: IBMC-CNRS, UPR 9002, Université Louis Pasteur, F-67084 Strasbourg Cedex, France.
authors.forename: V
authors.initials: V
authors.lastname: Oliéric
authors.forename: E
authors.initials: E
authors.lastname: Ennifar
authors.forename: A
authors.initials: A
authors.lastname: Meents
authors.forename: M
authors.initials: M
authors.lastname: Fleurant
authors.forename: C
authors.initials: C
authors.lastname: Besnard
authors.forename: P
authors.initials: P
authors.lastname: Pattison
authors.forename: M
authors.initials: M
authors.lastname: Schiltz
authors.forename: C
authors.initials: C
authors.lastname: Schulze-Briese
authors.forename: P
authors.initials: P
authors.lastname: Dumas
mesh_terms: D001966:Bromine / Q000737:chemistry / Q000528:radiation effects; D007700:Kinetics; D046911:Macromolecular Substances / Q000737:chemistry* / Q000528:radiation effects*; D011188:Potassium / Q000737:chemistry / Q000528:radiation effects; D012313:RNA / Q000737:chemistry; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013059:Spectrum Analysis, Raman; D014961:X-Ray Diffraction; D014965:X-Rays
chemical_list: D046911:Macromolecular Substances; D012313:RNA; D011188:Potassium; D001966:Bromine