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. 2012 Dec 14;288(5):3439–3448. doi: 10.1074/jbc.M112.431932

FIGURE 4.

FIGURE 4.

Equilibrium titration of MalE binding to MalFGK2 in nanodiscs. The MalFGK2 complex in nanodiscs was incubated with ATP (1 mm), BMOE (50 μm), and vanadate (10 μm) as indicated (10 min at room temperature). The nanodiscs were repurified on a desalting G25 column before fluorescence quenching assay using ATTO-655-labeled MalE (20 nm). Three independent measurements were performed, and the data points were fitted to a one-site binding equation (mean and standard deviation results are presented in Table 1). A, MalFGK2 wild type. B, MalFGK2 with mutations MalKS83C/MalKC40S (MalK83-MalK83). C, MalFGK2 with mutations MalFP396C/MalGD185C/MalKC40S (MalF396-MalG185). D, MalFGK2 with mutations MalFL446C/MalGS234C/MalKC40S (MalF446-MalG234). The left and right panels are the same curves but fitted to a different x axis.