Two-colour single-molecule photoinduced electron transfer fluorescence imaging microscopy of chaperone dynamics

Schubert, Jonathan, Schulze, Andrea, Prodromou, Chrisostomos and Neuweiler, Hannes (2021) Two-colour single-molecule photoinduced electron transfer fluorescence imaging microscopy of chaperone dynamics. Nature Communications, 12. a6964 1-12. ISSN 2041-1723

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Abstract

Many proteins are molecular machines, whose function is dependent on multiple conformational changes that are initiated and tightly controlled through biochemical stimuli. Their mechanistic understanding calls for spectroscopy that can probe simultaneously such structural coordinates. Here we present two-colour fluorescence microscopy in combination with photoinduced electron transfer (PET) probes as a method that simultaneously detects two structural coordinates in single protein molecules, one colour per coordinate. This contrasts with the commonly applied resonance energy transfer (FRET) technique that requires two colours per coordinate. We demonstrate the technique by directly and simultaneously observing three critical structural changes within the Hsp90 molecular chaperone machinery. Our results reveal synchronicity of conformational motions at remote sites during ATPase-driven closure of the Hsp90 molecular clamp, providing evidence for a cooperativity mechanism in the chaperone’s catalytic cycle. Single-molecule PET fluorescence microscopy opens up avenues in the multi-dimensional exploration of protein dynamics and allosteric mechanisms.

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 30 Nov 2021 09:57
Last Modified: 30 Nov 2021 10:00
URI: http://sro.sussex.ac.uk/id/eprint/103153

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