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. 2018 Apr 12;46(10):5159–5170. doi: 10.1093/nar/gky278

Figure 7.

Figure 7.

Equilibrium binding of TDGN140A, unmodified and sumoylated, to G·caC DNA (1 nM) monitored by fluorescence anisotropy. (A) Data for TDGN140A binding to G·caC DNA, fitted to a two-site model (using Dynafit), gives Kd = 2.3 ± 0.6 nM, Kd2 = 820 ± 500 nM, rD= 0.184 ± 0.0033, rED = 0.251 ± 0.004, and rEED = 0.284 ± 0.004. (B) Binding of TDGN140A∼SUMO-1 to G·caC DNA fitted to a two-site model (black line) gives Kd = 14 ± 2 nM, Kd2 = 24000 ± 16000 nM, rD= 0.189 ± 0.002, rED = 0.255 (fixed), and rEED = 0.285 (fixed). Fitting to a one site model (red line) yields essentially the same affinity, Kd = 16 ± 3 nM, with rD = 0.190 ± 0.002, and rED = 0.257 ± 0.002. (C) Binding of TDGN140A∼SUMO-2 to G·caC DNA fitted to a two-site model (black line) gives Kd = 13 ± 2 nM, Kd2 = 5000 ± 2000 nM, rD = 0.185 ± 0.003, rED = 0.251 (fixed), and rEED = 0.281 (fixed). Fitting to a one site model (red line) yields Kd = 20 ± 3 nM, with rD = 0.187 ± 0.002, and rED = 0.260 ± 0.002. Fitting for TDG∼SUMO-1 and -2 employed fixed values for rED and rEED, using the same approach described for fitting binding of these enzymes to G·fC DNA (Figure 6).