High-Resolution Protein Structure Determination by Serial Femtosecond Crystallography
Abstract Not Provided
CHEM↗
Engineering topics
Seibert, M. M.
Publications and source records attributed to Seibert, M. M..
Femtosecond X-ray Diffraction From Two-Dimensional Protein Crystals
Here we present femtosecond x-ray diffraction patterns from two-dimensional (2-D) protein crystals using an x-ray free electron laser (XFEL). To date it has not been possible to acquire x-ray diffraction from individual 2-D protein crystals due to radiation damage. However, the intense and ultrafast pulses generated by an XFEL permits a new method of collecting diffraction data before the sample is destroyed. Utilizing a diffract-before-destroy methodology at the Linac Coherent Light Source, we observed Bragg diffraction to better than 8.5 Å resolution for two different 2-D protein crystal samples that were maintained at room temperature. These proof-of-principle results show promise for structural analysis of both soluble and membrane proteins arranged as 2-D crystals without requiring cryogenic conditions or the formation of three-dimensional crystals.
36 MATERIALS SCIENCE↗
Single Particle Difraction at FLASH
Single-pulse coherent diffraction patterns have been collected from randomly injected single particles with a soft X-ray free-electron laser (FEL). The intense focused FEL pulse gives a high-resolution low-noise coherent diffraction pattern of the object before that object turns into a plasma and explodes. A diffraction pattern of a single particle will only be recorded when the particle arrival into the FEL interaction region coincides with FEL pulse arrival and detector integration. The properties of the experimental apparatus coinciding with these three events set the data acquisition rate. For our single particle FLASH diffraction imaging experiments: (1) an aerodynamic lens stack prepared a particle beam that consisted of particles moving at 150-200 m/s positioned randomly in space and time, (2) the 10 fs long FEL pulses were delivered at a fixed rate, and (3) the detector was set to integrate and readout once every two seconds. The effect of these experimental parameters on the rate of data acquisition using randomly injected particles will be discussed. Overall, the ultrashort FEL pulses do not set the limit of the data acquisition, more important is the effective interaction time of the particle crossing the FEL focus, the pulse sequence structure and the detector readout rate. Example diffraction patterns of randomly injected ellipsoidal iron oxide nanoparticles in different orientations are presented. This is the first single particle diffraction data set of identical particles in different orientations collected on a shot-to-shot basis. This data set will be used to test algorithms for recovering 3D structure from single particle diffraction.
36 MATERIALS SCIENCE↗