Time-resolved neutron scattering provides new insight into protein substrate processing by a AAA+ unfoldase. - Université Grenoble Alpes Accéder directement au contenu
Article Dans Une Revue Scientific Reports Année : 2017

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

Résumé

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.
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Dates et versions

hal-01465686 , version 1 (24-01-2018)

Identifiants

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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, 2017, 7, pp.40948. ⟨10.1038/srep40948⟩. ⟨hal-01465686⟩
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