Fig. 5.

Comparison of off-rates derived from analysis of bond lifetimes collected by the descriptive statistical and HMM methods. (A). Exponential bond lifetime distributions for the interaction of WT A1 and GC. An ensemble of ∼50 bond lifetimes, defined as the time span from association to dissociation of one bond, was pooled by the descriptive statistical method (blue squares). Another ensemble of ∼200 bond lifetimes was measured by the HMM method from the same raw data and pooled (red squares). For data obtained by each method, the natural log of the survival frequency with bond lifetimes >t_b was plotted against t_b and fitted by a straight line. The negative slopes of the best-fits represent the off-rate k_off. (B). Comparison of off-rates estimated by the descriptive statistical and HMM for WT, G1324S (Type 2M) and R1450E (Type 2B) A1s versus GC. The error bars show the 95% confidence interval for each method