Format

Send to

Choose Destination
Structure. 2018 Jun 5;26(6):848-856.e3. doi: 10.1016/j.str.2018.04.004. Epub 2018 May 10.

Atomic Resolution Cryo-EM Structure of β-Galactosidase.

Author information

1
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
2
Department of Electrical and Computer Engineering, Duke University, Durham, NC 27708, USA.
3
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
4
Laboratory of Biophysical Chemistry, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892, USA.
5
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA. Electronic address: subramas@mail.nih.gov.

Abstract

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.

KEYWORDS:

atomic resolution; computer-aided drug discovery; drift correction; drug discovery; high-resolution protein structure; precision medicine; radiation damage; single-particle cryo-EM

PMID:
29754826
PMCID:
PMC6129192
DOI:
10.1016/j.str.2018.04.004
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center