Invited Speaker 23rd International Society of Magnetic Resonance Conference 2023

Novel, affordable strategies for in-cell analysis of IDPs expressed in situ at 310K (#20)

Francois-Xavier Theillet 1 , Helene Cherot 2 3
  1. CNRS, Gif-sur-Yvette, France
  2. Paris-Saclay University, Saclay, France
  3. CEA-Saclay, Gif-sur-Yvette, France

"In-cell structural biology by NMR is appealing in many regards: It proposes, among others, to investigate conformational equilibria or ligand binding/processing in very relevant conditions, i.e in cells [1,2]. The approach comes with a number of challenges, among which i) the many difficulties in sample production, and ii) recording at 310K for a maximum relevance, which yields poor 1H-15N spectra for IDPs (because of water-amide proton exchange); iii) important signal losses due to promiscuous, transient interactions with cellular entities, which, in turn, urges to use (too) high concentrations of the studied proteins. We will show how we are trying to facilitate in-cell NMR studies, using new production methods in situ, new labeling schemes, and better adapted pulse sequences."

  1. Theillet F.-X. In-Cell Structural Biology by NMR: The Benefits of the Atomic-Scale. Chem. Rev. 122 (2022). https://doi.org/10.1021/acs.chemrev.1c00937
  2. Theillet F.X., Luchinat E. In-cell NMR: Why and how? Prog Nucl Reson Magn Spectr. 132-133 (2022). https://doi.org/10.1016/j.pnmrs.2022.04.002”