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Structure paper

TitleExperimental Phasing of MicroED Data Using Radiation Damage.
Journal, issue, pagesStructure, Vol. 28, Issue 4, Page 458-464.e2, Year 2020
Publish dateApr 7, 2020
AuthorsMichael W Martynowycz / Johan Hattne / Tamir Gonen /
PubMed AbstractWe previously demonstrated that microcrystal electron diffraction (MicroED) can be used to determine atomic-resolution structures from vanishingly small three-dimensional crystals. Here, we present ...We previously demonstrated that microcrystal electron diffraction (MicroED) can be used to determine atomic-resolution structures from vanishingly small three-dimensional crystals. Here, we present an example of an experimentally phased structure using only MicroED data. The structure of a seven-residue peptide is solved starting from differences to the diffraction intensities induced by structural changes due to radiation damage. The same wedge of reciprocal space was recorded twice by continuous-rotation MicroED from a set of 11 individual crystals. The data from the first pass were merged to make a "low-dose dataset." The data from the second pass were similarly merged to form a "damaged dataset." Differences between these two datasets were used to identify a single heavy-atom site from a Patterson difference map, and initial phases were generated. Finally, the structure was completed by iterative cycles of modeling and refinement.
External linksStructure / PubMed:32023481 / PubMed Central
MethodsEM (electron crystallography)
Resolution1.4 Å
Structure data

21202

6vhb

EMDB-21202, PDB-6vhb:
1.4A low-dose structure of GSNQNNF determined from initial phases generated using radiation damage
Method: EM (electron crystallography) / Resolution: 1.4 Å

21203

6vhc

EMDB-21203, PDB-6vhc:
1.4A damaged structure of GSNQNNF used to determine initial phases from radiation damage
Method: EM (electron crystallography)

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-ACT:
ACETATE ION

ChemComp-HOH:
WATER

Source
  • synthetic construct (others)
KeywordsPROTEIN FIBRIL / MicroED / damage / phasing / RIP

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