Figure 5.

Determination of dissociation rate (k_off) by DRaCALA.

(A) Representative DRaCALA images of protein-ligand spots taken at various time points after initiation of competition. The competition reaction was initiated by addition of 1 mM of unlabeled c-di-GMP to a mixture of ³²P-c-di-GMP and His-MBP-Alg44_PilZ. (B) F_B of ³²P-c-di-GMP is plotted against time after the addition of unlabeled competitor. k_off is calculated by nonlinear regression analysis fit to a one phase exponential decay. Adapted with permission from Roelofs et al. (2011).