National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35 GM128867
United States
Citation
Journal: IUCrJ / Year: 2020 Title: Atomic structures determined from digitally defined nanocrystalline regions. Authors: Marcus Gallagher-Jones / Karen C Bustillo / Colin Ophus / Logan S Richards / Jim Ciston / Sangho Lee / Andrew M Minor / Jose A Rodriguez / Abstract: Nanocrystallography has transformed our ability to interrogate the atomic structures of proteins, peptides, organic molecules and materials. By probing atomic level details in ordered sub-10 nm ...Nanocrystallography has transformed our ability to interrogate the atomic structures of proteins, peptides, organic molecules and materials. By probing atomic level details in ordered sub-10 nm regions of nanocrystals, scanning nanobeam electron diffraction extends the reach of nanocrystallography and in principle obviates the need for diffraction from large portions of one or more crystals. Scanning nanobeam electron diffraction is now applied to determine atomic structures from digitally defined regions of beam-sensitive peptide nanocrystals. Using a direct electron detector, thousands of sparse diffraction patterns over multiple orientations of a given crystal are recorded. Each pattern is assigned to a specific location on a single nanocrystal with axial, lateral and angular coordinates. This approach yields a collection of patterns that represent a tilt series across an angular wedge of reciprocal space: a scanning nanobeam diffraction tomogram. Using this diffraction tomogram, intensities can be digitally extracted from any desired region of a scan in real or diffraction space, exclusive of all other scanned points. Intensities from multiple regions of a crystal or from multiple crystals can be merged to increase data completeness and mitigate missing wedges. It is demonstrated that merged intensities from digitally defined regions of two crystals of a segment from the OsPYL/RCAR5 protein produce fragment-based solutions that can be refined to atomic resolution, analogous to structures determined by selected-area electron diffraction. In allowing atomic structures to now be determined from digitally outlined regions of a nanocrystal, scanning nanobeam diffraction tomography breaks new ground in nanocrystallography.
History
Deposition
Oct 15, 2019
Deposition site: RCSB / Processing site: RCSB
Revision 1.0
May 13, 2020
Provider: repository / Type: Initial release
Revision 1.1
Jun 30, 2021
Group: Data collection / Category: diffrn_detector / Item: _diffrn_detector.detector
Cryogen: NITROGEN Specimen holder model: GATAN 626 SINGLE TILT LIQUID NITROGEN CRYO TRANSFER HOLDER Temperature (max): 110 K / Temperature (min): 100 K
Image recording
Average exposure time: 3 sec. / Electron dose: 0.03 e/Å2 / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Num. of diffraction images: 100 / Num. of grids imaged: 1 / Num. of real images: 1
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