Quantitative Description of Intrinsically Disordered Proteins Using Single-Molecule FRET, NMR, and SAXS - Université Grenoble Alpes Accéder directement au contenu
Article Dans Une Revue Journal of the American Chemical Society Année : 2021

Quantitative Description of Intrinsically Disordered Proteins Using Single-Molecule FRET, NMR, and SAXS

Samuel Naudi-Fabra
Martin Blackledge
Sigrid Milles

Résumé

Studying the conformational landscape of intrinsically disordered and partially folded proteins is challenging and only accessible to a few solution state techniques, such as nuclear magnetic resonance (NMR), small-angle scattering techniques, and single-molecule Förster resonance energy transfer (smFRET). While each of the techniques is sensitive to different properties of the disordered chain, such as local structural propensities, overall dimension, or intermediate- and long-range contacts, conformational ensembles describing intrinsically disordered proteins (IDPs) accurately should ideally respect all of these properties. Here we develop an integrated approach using a large set of FRET efficiencies and fluorescence lifetimes, NMR chemical shifts, and paramagnetic relaxation enhancements (PREs), as well as small-angle X-ray scattering (SAXS) to derive quantitative conformational ensembles in agreement with all parameters. Our approach is tested using simulated data (five sets of PREs and 15 FRET efficiencies) and validated experimentally on the example of the disordered domain of measles virus phosphoprotein, providing new insights into the conformational landscape of this viral protein that comprises transient structural elements and is more compact than an unfolded chain throughout its length. Rigorous cross-validation using FRET efficiencies, fluorescence lifetimes, and SAXS demonstrates the predictive nature of the calculated conformational ensembles and underlines the potential of this strategy in integrative dynamic structural biology.
Fichier principal
Vignette du fichier
Naudi Fabra et al. JACS.pdf (2.66 Mo) Télécharger le fichier
Origine Accord explicite pour ce dépôt

Dates et versions

hal-03549077 , version 1 (10-10-2022)

Identifiants

Citer

Samuel Naudi-Fabra, Maud Tengo, Malene Ringkjøbing Jensen, Martin Blackledge, Sigrid Milles. Quantitative Description of Intrinsically Disordered Proteins Using Single-Molecule FRET, NMR, and SAXS. Journal of the American Chemical Society, 2021, 143 (48), pp.20109-20121. ⟨10.1021/jacs.1c06264⟩. ⟨hal-03549077⟩
50 Consultations
127 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More