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. Author manuscript; available in PMC: 2015 Mar 15.
Published in final edited form as: Methods. 2013 Jul 23;66(2):168–179. doi: 10.1016/j.ymeth.2013.07.026

Figure 1.

Figure 1

Figure 1

Schematic of the experimental design. (A) Structural model of mouse Pgp crystallized in the inward facing, nucleotide-free conformation (3g5u). (B) Homology model of the two NBDs of Pgp in the outward facing conformation. For both conformations, the positions of T492 (green) and S1137 (red) are indicated in space fill. Modeling for panel (B) was completed using the outward facing conformation of Sav1866 (2hyd) as described [19]. (C) Confocal detection of freely diffusing FRETlabeled Pgp reconstituted in a liposome. Two laser foci are aligned for duty-cycle optimized alternating laser excitation with 488 nm and 594 nm. (D) Optical setup with two lasers triggered by an arbitrary waveform generator and synchronized TCSPC electronics for time-resolved single-molecule FRET recording.