Time-resolved neutron scattering provides new insight into protein substrate processing by a AAA+ unfoldase.

Abstract : We present a combination of small-angle neutron scattering, deuterium labelling and contrast variation, temperature activation and fluorescence spectroscopy as a novel approach to obtain time-resolved, structural data individually from macromolecular complexes and their substrates during active biochemical reactions. The approach allowed us to monitor the mechanical unfolding of a green fluorescent protein model substrate by the archaeal AAA+ PAN unfoldase on the sub-minute time scale. Concomitant with the unfolding of its substrate, the PAN complex underwent an energy-dependent transition from a relaxed to a contracted conformation, followed by a slower expansion to its initial state at the end of the reaction. The results support a model in which AAA ATPases unfold their substrates in a reversible power stroke mechanism involving several subunits and demonstrate the general utility of this time-resolved approach for studying the structural molecular kinetics of multiple protein remodelling complexes and their substrates on the sub-minute time scale.
Type de document :
Article dans une revue
Scientific Reports, Nature Publishing Group, 2017, 7, pp.40948. 〈10.1038/srep40948〉
Liste complète des métadonnées

Littérature citée [44 références]  Voir  Masquer  Télécharger

http://hal.univ-grenoble-alpes.fr/hal-01465686
Contributeur : Frank Thomas <>
Soumis le : mercredi 24 janvier 2018 - 13:46:18
Dernière modification le : lundi 19 février 2018 - 14:34:04

Fichier

Scientific_Reports_7_p40948.pd...
Fichiers éditeurs autorisés sur une archive ouverte

Identifiants

Collections

Citation

Ziad Ibrahim, Anne Martel, Martine Moulin, Henry S Kim, Michael Härtlein, et al.. Time-resolved neutron scattering provides new insight into protein substrate processing by a AAA+ unfoldase.. Scientific Reports, Nature Publishing Group, 2017, 7, pp.40948. 〈10.1038/srep40948〉. 〈hal-01465686〉

Partager

Métriques

Consultations de la notice

183

Téléchargements de fichiers

25