Kinetic and thermodynamic parameters of the GF14ε and GF14ω interaction with the H+-ATPase.

The kinetic parameters, association rate constants (kon) and dissociation rate constants (koff), were determined using the BIAcoreX system for SPR detection. Biotinylated peptide bL15Vp (600 resonance units) was captured on an Biacore SA sensor chip precoated with streptavidin. GF14ε or GF14ω were applied in the concentration ranges of 10 µM–7.8 nM in the absence or presence of 10 µM FC at 25°C, with a flow rate of 30 µl/min. Dissociation was monitored for five minutes. The data were evaluated using BIAevaluation 3.1 (Biacore AB). To obtain KD and Rmax values, a 1∶1 Langmuir binding model was fitted to the sensorgrams.