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. 2010 Oct 6;99(7):2125–2133. doi: 10.1016/j.bpj.2010.07.047

Figure 1.

Figure 1

(A) Representative binding curve for association of fluorescein-labeled NusDHFR with POPC/MTX-PE/RhoPE (99.5:0.5:0.02 molar proportions) vesicles, determined using the ultracentrifugation-based assay described in Materials and Methods. Data are fit to an equation of the form (% Bound) = A+B∗[MTX-PE]ext/([MTX-PE]ext + Kd), where [MTX-PE]ext is the concentration of MTX-PE exposed at the vesicles' outer surface, and A, B and Kd are adjustable parameters. (B and C) Dissociation constants determined for binding of DHFR (B) or NusDHFR (C) to vesicles prepared from POPC, 0.5 mol % MTX-PE, 0.02 mol % RhoPE, and the indicated molar percentages of POPE-PEG5000 (solid circles) or POPG (open circles). The Kd values shown (normalized to the value of Kd determined in the same experiment for vesicles lacking POPE-PEG5000/POPG) represent the mean (± SE) of values determined in three independent experiments. Data are fitted to theoretical curves as described in the text.