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

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

SUBMITTER: Schubert J 

PROVIDER: S-EPMC8630005 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature