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Increasing fluorescence transient observed on rapid mixing of [L91D,L-O]RBD/H (0.5 μM) with H-Ras.mantGppNHp (2 μM) and transient observed for the rapid mixing of [L91W]RBD/H (0.5 μM) with H-Ras.mantGppNHp (2 μM) at 25°C under the same conditions. The excitation wavelength was 313 nm with detection through a 360-nm cutoff filter. The fitted curve of the increasing transient corresponds to a pseudo-first-order rate constant of 90 s−1.
