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. 2020 Mar 10;295(17):5614–5625. doi: 10.1074/jbc.RA120.012588

Table 1.

Parameters for SC(1–246)-BODIPY and unlabeled WT SC(1–246) binding to four separate ProTQQQ preparations

Reference titrations of SC(1–246)-BODIPY with four separate ProTQQQ batches were obtained at two fixed probe concentrations. The competitive binding data for the ProTQQQ preparations were obtained by titrations of fixed concentrations of SC(1–246)-BODIPY probe, and SC(1–246) as competitor, with ProTQQQ. Data were fit simultaneously by the cubic equation to obtain the dissociation constant for ProTQQQ and SC(1–246)-BODIPY (KO, probe) and the competitor SC(1–246) (KC, competitor); the stoichiometric factor for SC(1–246)-BODIPY (n) and SC(1–246) (m); and the maximum fluorescence intensity (ΔFmax/Fo). Experimental error represents ± 2 S.D. Competitive equilibrium binding studies and data analysis were performed as described under “Experimental procedures.” SF, stoichiometric factor.

SC(1–246) SF (n or m) KO or KC ΔFmax/Fo
nm
SC(1–246)-BODIPY (n) 0.90 ± 0.03 2.9 ± 0.7 0.60 ± 0.01
SC(1–246) (m) 1.10 ± 0.05 0.7 ± 0.2
SC(1–246)-BODIPY (n) 0.90 ± 0.02 5.9 ± 0.8 0.59 ± 0.01
SC(1–246) (m) 1.10 ± 0.06 1.0 ± 0.3
SC(1–246)-BODIPY (n) 1.00 ± 0.04 4.1 ± 1.1 0.57 ± 0.01
SC(1–246) (m) 1.10 ± 0.05 0.7 ± 0.2
SC(1–246)-BODIPY (n) 1.01 ± 0.06 3.0 ± 1.4 0.58 ± 0.01
SC(1–246) (m) 1.20 ± 0.08 0.6 ± 0.4