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. 2017 Oct 30;114(46):12178–12183. doi: 10.1073/pnas.1712882114

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Fig. 1. — The 2B subdomain of UvrD populates at least four discrete rotational conformational states. (A) The 2B subdomain (blue) in the open and closed states in crystal structures. Movement of the 2B domain is monitored by the change in FRET efficiency between a donor (Cy3) and an acceptor (Cy5) on the 1B (A100C) and the 2B (A473C) subdomains. (B) A single-molecule time trace [Cy3 (green), Cy5 (red), and FRET efficiency (blue)] in imaging buffer plus 60 mM NaCl (25.0 °C) shows anticorrelated changes in Cy3 and Cy5 with a constant total intensity (black). Transitions are observed among four FRET states (S1 to S4) as shown by the hidden Markov fit (solid black line) to the trajectory. (C) Schematic of the four FRET substates and their interconversion rates obtained from hidden Markov analyses.