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We describe the deposition of four datasets consisting of X-ray diffraction images acquired using serial femtosecond crystallography experiments on microcrystals of human G protein-coupled receptors, grown and delivered in lipidic cubic phase, at the Linac Coherent Light Source. The receptors are: the human serotonin receptor 2B in complex with an agonist ergotamine, the human δ-opioid receptor in complex with a bi-functional peptide ligand DIPP-NH2, the human smoothened receptor in complex with an antagonist cyclopamine, and finally the human angiotensin II type 1 receptor in complex with the selective antagonist ZD7155. All four datasets have been deposited, with minimal processing, in an HDF5-based file format, which can be used directly for crystallographic processing with CrystFEL or other software. We have provided processing scripts and supporting files for recent versions of CrystFEL, which can be used to validate the data.
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To achieve a sensing diameter of 1-2 nanometers, the diatom shells were used as substrates to perform ion-channel reconstitution experiments. The immobilized diatom shell was functionalized using silane chemistry and lipid bilayer membranes were formed. Functionalization of the diatom shell surface improves bilayer formation probability from 1 out of 10 to 10 out of 10 as monitored by impedance spectroscopy. Self-insertion of outer membrane protein OmpF of E.Coli into the lipid membranes could be confirmed using single channel recordings, indicating that nano-BLMs had formed which allow for fully functional porin activity. The results indicate that biogenic silica nanoporous substrates can be simulated using a simplified two dimensional geometry to predict the current when a nanoparticle translocates through a single aperture. With their tiered three-dimensional structure, diatom shells can be used in to form nano-lipid bilayer membranes and can be used in ion-channel reconstitution experiments similar to synthetic nanoporous membranes.
The ASU School of Dance presents Undergraduate Projects Showing, October 25-26, with works by undergraduate dance students, performed at Margaret Gisolo Dance Studio.