16-17 March 2020
ILL4
Europe/Paris timezone

Integrated NMR and cryo-EM atomic-resolution structure determination of a half-megadalton enzyme complex

Not scheduled
15m
ILL4-rdc-1 - Amphi Chadwick (ILL4)

ILL4-rdc-1 - Amphi Chadwick

ILL4

110
Oral Session 8

Speaker

Adrien Favier (IBS CNRS)

Description

Atomic-resolution structure determination is the key requirement 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 approach1 that simultaneously uses NMR and EM data to overcome the limits of each of these methods. The approach enabled determination of the high-resolution structure of the 468 kDa large dodecameric aminopeptidase TET2 to a precision and accuracy below 1 Angstrom 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 specifically labelled methyl groups, and a 4.1 Angstrom 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 (up to 8 Angstroms) cryo-EM data are available, thus paving avenues for the structure determination of challenging biological assemblies.

Primary authors

Adrien Favier (IBS CNRS) Dr. Diego F. Gauto (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France) Leandro Farias Estrozi (IBS) Dr. Charles D. Schwieters (Center for Information Technology, National Institutes of Health, Bethesda, USA) Mr. Gregory Effantin (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France) Pavel Macek (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France; NMR-Bio, Grenoble, France) Dr. Remy Sounier (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France; Institut de Génomique Fonctionnelle, University of Montpellier, Montpellier, France) Astrid C. Sivertsen (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France) Elena Schmidt (Institute of Biophysical Chemistry, Goethe University Frankfurt am Main, Frankfurt, Germany) Rime Kerfah (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France; Graduate School of Science, Tokyo Metropolitan University, Tokyo, Japan) Guillaume Mas (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France; Biozentrum University of Basel, Basel, Switzerland) Jacques-Philippe Colletier Peter Guntert (Institute of Biophysical Chemistry, Goethe University Frankfurt am Main, Frankfurt, Germany; Laboratory of Physical Chemistry, ETH Zurich, Zurich, Switzerland; Graduate School of Science, Tokyo Metropolitan University, Tokyo, Japan) Dr. Guy Schoehn (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France) Jerome Boisbouvier (Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, France) Paul Schanda (IBS Grenoble)

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