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. 2012 Dec 6;288(6):4378–4388. doi: 10.1074/jbc.M112.388629

FIGURE 5.

FIGURE 5.

Inter-Kir6.2 subunit FRET efficiencies. A and B, FRET efficiency between CFP and YFP fused at N- or C-terminal of full-length Kir6.2 or transmembrane domains. Apparent FRET efficiencies obtained with different measured CFP/YFP ratios were fitted with equations derived from tetrameric FRET model based on FRET rate theory (see text); FRET efficiency between 6.2C/6.2Y, 6.2tmC/6.2tmY, and C6.2/Y6.2 (24.5, 28.9, and 0.3%), indicate close proximity of C termini, but not N termini; C6.2/6.2Y, but not Y6.2/6.2C indicates proximity of C and N termini, but with clear anisotropy; only C-terminal CFP can transfer energy to N-terminal YFP. C, FRET efficiencies of CFP/YFP tags at N or C termini of Kir6.2 subunits, calculated using the tetrameric FRET model described by Cheng et al. (24). D, FRET efficiencies of CFP/YFP fused at N or C termini of truncated Kir6.2 transmembrane domain.