Steady-state fluorescence anisotropy measurement of TRPC6_(801–878) WT and selected mutants to fluorescently labeled S100A1 protein.

<p>DNS- S100A1 protein (232 µM) was titrated with TRPC6 fusion protein and the Fb was calculated using equation 1 as was described in material and methods.Binding isotherms and dissociatin constants were calculated by fitting the data to the equation 2 as was described in material and methods. Values are expressed as the mean ± standard deviation (SD) measured from at least from three independent experiments. Binding isotherms of wild-type TRPC6 _(801–878) is represented as black circles, single mutant is TRPC6_(801–878) R864A as white circles and triple mutant TRPC6_(801–878) K859A/R860A/R864A as black squares.