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TitleAtomic Resolution Cryo-EM Structure of β-Galactosidase.
Journal, issue, pagesStructure, Vol. 26, Issue 6, Page 848-856.e3, Year 2018
Publish dateJun 5, 2018
AuthorsAlberto Bartesaghi / Cecilia Aguerrebere / Veronica Falconieri / Soojay Banerjee / Lesley A Earl / Xing Zhu / Nikolaus Grigorieff / Jacqueline L S Milne / Guillermo Sapiro / Xiongwu Wu / Sriram Subramaniam /
PubMed AbstractThe advent of direct electron detectors has enabled the routine use of single-particle cryo-electron microscopy (EM) approaches to determine structures of a variety of protein complexes at near- ...The advent of direct electron detectors has enabled the routine use of single-particle cryo-electron microscopy (EM) approaches to determine structures of a variety of protein complexes at near-atomic resolution. Here, we report the development of methods to account for local variations in defocus and beam-induced drift, and the implementation of a data-driven dose compensation scheme that significantly improves the extraction of high-resolution information recorded during exposure of the specimen to the electron beam. These advances enable determination of a cryo-EM density map for β-galactosidase bound to the inhibitor phenylethyl β-D-thiogalactopyranoside where the ordered regions are resolved at a level of detail seen in X-ray maps at ∼ 1.5 Å resolution. Using this density map in conjunction with constrained molecular dynamics simulations provides a measure of the local flexibility of the non-covalently bound inhibitor and offers further opportunities for structure-guided inhibitor design.
External linksStructure / PubMed:29754826 / PubMed Central
MethodsEM (single particle)
Resolution1.9 Å
Structure data

EMDB-7770, PDB-6cvm:
Atomic resolution cryo-EM structure of beta-galactosidase
Method: EM (single particle) / Resolution: 1.9 Å

Chemicals

ChemComp-PTQ:
2-phenylethyl 1-thio-beta-D-galactopyranoside

ChemComp-MG:
Unknown entry

ChemComp-NA:
Unknown entry

ChemComp-HOH:
WATER

Source
  • Escherichia coli K-12 (bacteria)
  • escherichia coli (strain k12) (bacteria)
KeywordsHYDROLASE / drift correction / radiation damage / drug discovery / precision medicine / computer-aided drug discovery

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