Atomic Resolution Cryo-EM Structure of β-Galactosidase.

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Bartesaghi, A; Aguerrebere, C; Falconieri, V; Banerjee, S; Earl, LA; Zhu, X; Grigorieff, N; Milne, JLS; Sapiro, G; Wu, X; Subramaniam, S

Published Date

  • June 2018

Published In

Volume / Issue

  • 26 / 6

Start / End Page

  • 848 - 856.e3

PubMed ID

  • 29754826

Electronic International Standard Serial Number (EISSN)

  • 1878-4186

International Standard Serial Number (ISSN)

  • 0969-2126

Digital Object Identifier (DOI)

  • 10.1016/j.str.2018.04.004

Language

  • eng