Breaking Cryo-EM Resolution Barriers to Facilitate Drug Discovery.

Published

Journal Article

Recent advances in single-particle cryoelecton microscopy (cryo-EM) are enabling generation of numerous near-atomic resolution structures for well-ordered protein complexes with sizes ≥ ∼200 kDa. Whether cryo-EM methods are equally useful for high-resolution structural analysis of smaller, dynamic protein complexes such as those involved in cellular metabolism remains an important question. Here, we present 3.8 Å resolution cryo-EM structures of the cancer target isocitrate dehydrogenase (93 kDa) and identify the nature of conformational changes induced by binding of the allosteric small-molecule inhibitor ML309. We also report 2.8-Å- and 1.8-Å-resolution structures of lactate dehydrogenase (145 kDa) and glutamate dehydrogenase (334 kDa), respectively. With these results, two perceived barriers in single-particle cryo-EM are overcome: (1) crossing 2 Å resolution and (2) obtaining structures of proteins with sizes < 100 kDa, demonstrating that cryo-EM can be used to investigate a broad spectrum of drug-target interactions and dynamic conformational states.

Full Text

Duke Authors

Cited Authors

  • Merk, A; Bartesaghi, A; Banerjee, S; Falconieri, V; Rao, P; Davis, MI; Pragani, R; Boxer, MB; Earl, LA; Milne, JLS; Subramaniam, S

Published Date

  • June 2016

Published In

Volume / Issue

  • 165 / 7

Start / End Page

  • 1698 - 1707

PubMed ID

  • 27238019

Pubmed Central ID

  • 27238019

Electronic International Standard Serial Number (EISSN)

  • 1097-4172

International Standard Serial Number (ISSN)

  • 0092-8674

Digital Object Identifier (DOI)

  • 10.1016/j.cell.2016.05.040

Language

  • eng