Macromolecular interactions in vitro, comparing classical and novel approaches - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue European Biophysics Journal Année : 2021

Macromolecular interactions in vitro, comparing classical and novel approaches

Guillaume Bec
  • Fonction : Auteur
  • PersonId : 1098574

Résumé

Biophysical quantification of protein interactions is central to unveil the molecular mechanisms of cellular processes. Researchers can choose from a wide panel of biophysical methods that quantify molecular interactions in different ways, including both classical and more novel techniques. We report the outcome of an ARBRE-MOBIEU training school held in June 2019 in Gif-sur-Yvette, France (https://mosbio.sciencesconf.org/). Twenty European students benefited from a week’s training with theoretical and practical sessions in six complementary approaches: (1) analytical ultracentrifugation with or without a fluorescence detector system (AUC-FDS), (2) isothermal titration calorimetry (ITC), (3) size exclusion chromatography coupled to multi-angle light scattering (SEC-MALS), (4) bio-layer interferometry (BLI), (5) microscale thermophoresis (MST) and, (6) switchSENSE. They implemented all these methods on two examples of macromolecular interactions with nanomolar affinity: first, a protein–protein interaction between an artificial alphaRep binder, and its target protein, also an alphaRep; second, a protein-DNA interaction between a DNA repair complex, Ku70/Ku80 (hereafter called Ku), and its cognate DNA ligand. We report the approaches used to analyze the two systems under study and thereby showcase application of each of the six techniques. The workshop provided students with improved understanding of the advantages and limitations of different methods, enabling future choices concerning approaches that are most relevant or informative for specific kinds of sample and interaction.
Fichier principal
Vignette du fichier
islandora_117196.pdf (7.66 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-03224764 , version 1 (11-05-2021)

Identifiants

Citer

Christophe Velours, Magali Aumont-Nicaise, Stephan Uebel, Patrick England, Adrian Velazquez-Campoy, et al.. Macromolecular interactions in vitro, comparing classical and novel approaches. European Biophysics Journal, 2021, 50, pp.313-330. ⟨10.1007/s00249-021-01517-5⟩. ⟨hal-03224764⟩
139 Consultations
573 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More