Fig. 5.

Kinetic and equilibrium information for CaM₁₂ (left) and CaM₃₄ (Right) at varying calcium concentrations. (A, B) Zero-force extrapolated unfolding rate constants. (C, D) Zero-force extrapolated folding rate constants. (E, F) Equilibrium free energies of folding. Dashed lines in (C) and (D) are the calculated diffusion-limited on-rates for one calcium ion assuming a rate limit of 10⁸ M^-1 s^-1. Filled circles represent averages over several molecules at a particular concentration. Empty circles in (B, D, F) are values for the short-lived state as shown in Fig. 4, Middle. Error bars represent the SEM. Colored continuous lines are fits to the data. Vertical grey lines in (A–D) represent the value of K_TS1 as obtained from the fit. The shaded areas are the concentration range below , where the protein is predominantly ligand-free.