Upgrade of the small-angle X-ray scattering beamline X33 at the European Molecular Biology Laboratory, Hamburg

Abstract

The small-angle X-ray scattering beamline X33 of the European Molecular Biology Laboratory (EMBL) at the DORIS III storage ring [Deutsches Elektronen Synchrotron (DESY) Hamburg] was used for more than two decades to study the structure of non-crystalline biological systems. During recent years the beamline's scope has changed and is now predominantly used to analyze solutions of biological macromolecules. Owing to renewed interest in solution scattering studies from the biological community, the workload on the beamline has steadily increased. A major upgrade of X33 was performed to improve the beamline stability and data quality, to shorten the measurement time and to ensure user-friendly operation. The upgrade involved all major components of the beamline, including the optical system (monochromator, mirror, slits, beam monitors), electronics, control and acquisition software, X-ray detector system and the sample environment. The upgrade improved the brilliance by a factor of about three and the measuring time was reduced by a factor of seven. The knowledge and experience gained during the implementation of the upgrades to X33, may aid the design process for the BioSAXS beamline to be constructed for the PETRA-3 facility at DESY.

Characterization of Enzymes from Legionella pneumophila Involved in Reversible Adenylylation of Rab1 Protein

“…Purification of SidD 36 -507 (18). SAXS Measurements and Modeling-Small angle x-ray scattering (SAXS) data were collected at the European Molecular Biology Laboratory (EMBL) X33 beamline on the storage ring DORIS III (Deutsches Elektronen-Synchrotron (DESY)) (25). Protein buffer consisted of 20 mM Hepes, pH 8.0, 50 mM NaCl, 1 mM MgCl 2 , 2 mM dithioerythritol.…”

Structural Basis for the 14-3-3 Protein-dependent Inhibition of the Regulator of G Protein Signaling 3 (RGS3) Function

“…No measurable radiation damage was detected by comparison of eight successive time frames with 15-s exposures. The data were averaged after normalization to the intensity of the transmitted beam, and the scattering of the buffer was subtracted using PRIMUS (33). The forward scattering I(0) and the radius of gyration R g were evaluated using the Guinier approximation (34).…”

The Role of Hydration in Protein Stability: Comparison of the Cold and Heat Unfolded States of Yfh1

“…34 Previously desalted solutions of Yfh1 in 20 mM Hepes (pH 7) at 5.0 mg/ml were measured at 0, 10, 20, 40 and 50°C on different aliquots from the same protein batch. Before the measurements, 2 mM DTT was added to the samples to reduce radiation damage.…”