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Title | Integrated NMR and cryo-EM atomic-resolution structure determination of a half-megadalton enzyme complex. |
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Journal, issue, pages | Nat Commun, Vol. 10, Issue 1, Page 2697, Year 2019 |
Publish date | Jun 19, 2019 |
Authors | Diego F Gauto / Leandro F Estrozi / Charles D Schwieters / Gregory Effantin / Pavel Macek / Remy Sounier / Astrid C Sivertsen / Elena Schmidt / Rime Kerfah / Guillaume Mas / Jacques-Philippe Colletier / Peter Güntert / Adrien Favier / Guy Schoehn / Paul Schanda / Jerome Boisbouvier / |
PubMed Abstract | Atomic-resolution structure determination is crucial for understanding protein function. Cryo-EM and NMR spectroscopy both provide structural information, but currently cryo-EM does not routinely ...Atomic-resolution structure determination is crucial for understanding protein function. Cryo-EM and NMR spectroscopy both provide structural information, but currently cryo-EM does not routinely give access to atomic-level structural data, and, generally, NMR structure determination is restricted to small (<30 kDa) proteins. We introduce an integrated structure determination approach that simultaneously uses NMR and EM data to overcome the limits of each of these methods. The approach enables structure determination of the 468 kDa large dodecameric aminopeptidase TET2 to a precision and accuracy below 1 Å by combining secondary-structure information obtained from near-complete magic-angle-spinning NMR assignments of the 39 kDa-large subunits, distance restraints from backbone amides and ILV methyl groups, and a 4.1 Å resolution EM map. The resulting structure exceeds current standards of NMR and EM structure determination in terms of molecular weight and precision. Importantly, the approach is successful even in cases where only medium-resolution cryo-EM data are available. |
External links | Nat Commun / PubMed:31217444 / PubMed Central |
Methods | EM (single particle) / NMR (solution) / NMR (solid-state) |
Resolution | 4.1 Å |
Structure data | EMDB-4179, PDB-6f3k: |
Chemicals | ChemComp-ZN: |
Source |
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Keywords | PEPTIDE BINDING PROTEIN / peptidase / protein quality control / oligomer / aminopeptidase |