title: Measurements of accurate x-ray scattering data of protein solutions using small stationary sample cells.
abstract: In this paper, we report a method of precise in situ x-ray scattering measurements on protein solutions using small stationary sample cells. Although reduction in the radiation damage induced by intense synchrotron radiation sources is indispensable for the correct interpretation of scattering data, there is still a lack of effective methods to overcome radiation-induced aggregation and extract scattering profiles free from chemical or structural damage. It is found that radiation-induced aggregation mainly begins on the surface of the sample cell and grows along the beam path; the diameter of the damaged region is comparable to the x-ray beam size. Radiation-induced aggregation can be effectively avoided by using a two-dimensional scan (2D mode), with an interval as small as 1.5 times the beam size, at low temperature (e.g., 4 degrees C). A radiation sensitive protein, bovine hemoglobin, was used to test the method. A standard deviation of less than 5% in the small angle region was observed from a series of nine spectra recorded in 2D mode, in contrast to the intensity variation seen using the conventional stationary technique, which can exceed 100%. Wide-angle x-ray scattering data were collected at a standard macromolecular diffraction station using the same data collection protocol and showed a good signal/noise ratio (better than the reported data on the same protein using a flow cell). The results indicate that this method is an effective approach for obtaining precise measurements of protein solution scattering.
authors.affiliation: MacCHESS, Cornell High Energy Synchrotron Source, Cornell University, Ithaca, New York 14853, USA. xh48@cornell.edu
authors.forename: Xinguo
authors.initials: X
authors.lastname: Hong
authors.forename: Quan
authors.initials: Q
authors.lastname: Hao
mesh_terms: D000818:Animals; D002417:Cattle; D006454:Hemoglobins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D014961:X-Ray Diffraction / Q000295:instrumentation* / Q000379:methods*
chemical_list: D006454:Hemoglobins