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TitleAtomic structures determined from digitally defined nanocrystalline regions.
Journal, issue, pagesIUCrJ, Vol. 7, Issue Pt 3, Page 490-499, Year 2020
Publish dateMay 1, 2020
AuthorsMarcus Gallagher-Jones / Karen C Bustillo / Colin Ophus / Logan S Richards / Jim Ciston / Sangho Lee / Andrew M Minor / Jose A Rodriguez /
PubMed AbstractNanocrystallography 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.
External linksIUCrJ / PubMed:32431832 / PubMed Central
MethodsEM (electron crystallography)
Resolution0.9 - 1.351 Å
Structure data

PDB-6uop:
OsPYL/RCAR5 (24 - 29) solved by nanobeam diffraction tomography
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 1.351 Å

PDB-6uoq:
OsPYL/RCAR5 residues 24-29 solved from electron diffraction stills
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 1.007 Å

PDB-6uor:
MicroED structure of OsPYL/RCAR5 (24-29) at 3 e-/A^2
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 0.9 Å

PDB-6uos:
MicroED structure of OsPYL/RCAR5 (24-29) at 6 e-/A^2
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 0.9 Å

PDB-6uou:
MicroED structure of OsPYL/RCAR5 (24-29) at 9 e-/A^2
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 1.04 Å

PDB-6uow:
MicroED structure of OsPYL/RCAR5 (24-29) at 12 e-/A^2
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 1.2 Å

Source
  • oryza sativa (Asian cultivated rice)
KeywordsPROTEIN FIBRIL / protofilament

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