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Title | Breaking Cryo-EM Resolution Barriers to Facilitate Drug Discovery. |
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Journal, issue, pages | Cell, Vol. 165, Issue 7, Page 1698-1707, Year 2016 |
Publish date | Jun 16, 2016 |
Authors | Alan Merk / Alberto Bartesaghi / Soojay Banerjee / Veronica Falconieri / Prashant Rao / Mindy I Davis / Rajan Pragani / Matthew B Boxer / Lesley A Earl / Jacqueline L S Milne / Sriram Subramaniam / |
PubMed Abstract | 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. ...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. |
External links | Cell / PubMed:27238019 / PubMed Central |
Methods | EM (single particle) |
Resolution | 1.8 - 3.8 Å |
Structure data | EMDB-8191: Cryo-EM structure of lactate dehydrogenase (LDH) in complex with GSK2837808A EMDB-8192, PDB-5k10: |
Chemicals | ChemComp-HOH: ChemComp-NDP: |
Source |
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Keywords | OXIDOREDUCTASE / lactate dehydrogenase / small metabolic complex / small molecule inhibitor / isocitrate dehydrogenase / glutamate dehydrogenase |