Open AccessHigh‐resolution wide‐angle X‐ray scattering of protein solutions: effect of beam dose on protein integrity
Author(s) -
Fischetti Robert F.,
Rodi Diane J.,
Mirza Ahmed,
Irving Thomas C.,
Kondrashkina Elena,
Makowski Lee
Publication year - 2003
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s0909049503016583
Subject(s) - beamline , scattering , small angle x ray scattering , synchrotron , protein structure , crystallography , chemistry , biological small angle scattering , protein folding , physics , beam (structure) , optics , small angle neutron scattering , biochemistry , neutron scattering
Wide‐angle X‐ray scattering patterns from proteins in solution contain information relevant to the determination of protein fold. At relevant scattering angles, however, these data are weak, and the degree to which they might be used to categorize the fold of a protein is unknown. Preliminary work has been performed at the BioCAT insertion‐device beamline at the Advanced Photon Source which demonstrates that one can collect X‐ray scattering data from proteins in solution to spacings of at least 2.2 Å ( q = 2.8 Å −1 ). These data are sensitive to protein conformational states, and are in good agreement with the scattering predicted by the program CRYSOL using the known three‐dimensional atomic coordinates of the protein. An important issue in the exploitation of this technique as a tool for structural genomics is the extent to which the high intensity of X‐rays available at third‐generation synchrotron sources chemically or structurally damage proteins. Various data‐collection protocols have been investigated demonstrating conditions under which structural degradation of even sensitive proteins can be minimized, making this technique a viable tool for protein fold categorization, the study of protein folding, unfolding, protein–ligand interactions and domain movement.