High-resolution wide-angle X-ray scattering of protein solutions: effect of beam dose on protein integrity.
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 A (q = 2.8 A(-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.
Fischetti
Robert F
RF
GM/CA-CAT, Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439, USA.
Rodi
Diane J
DJ
Mirza
Ahmed
A
Irving
Thomas C
TC
Kondrashkina
Elena
E
Makowski
Lee
L
D000818:Animals; D016477:Artifacts; D002417:Cattle; D003198:Computer Simulation; D003574:Cytochrome c Group / Q000737:chemistry / Q000528:radiation effects; D004355:Drug Stability; D004912:Erythrocytes / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry / Q000528:radiation effects; D006736:Horses; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry / Q000528:radiation effects; D011487:Protein Conformation; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry* / Q000528:radiation effects*; D012542:Scattering, Radiation; D012996:Solutions / Q000737:chemistry / Q000528:radiation effects; D014961:X-Ray Diffraction / Q000379:methods*; D014965:X-Rays
D003574:Cytochrome c Group; D006454:Hemoglobins; D009211:Myoglobin; D011506:Proteins; D012996:Solutions