PMC:2724026 / 33191-33860 JSONTXT

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Kinetics For WT and mutants, refolding and unfolding were monitored by changes in fluorescence, using an Applied Photophysics SX.18MV stopped-flow fluorimeter. An excitation wavelength of 280 nm was used with a 320 nm cut-off filter; the final concentration of protein was 1–2 μM. WT kinetics were also monitored by CD using an Applied Photophysics Π⁎-180 instrument, with a final maximum concentration of protein of 5 μM. In both cases, the stopped-flow apparatus was maintained at 25(± 0.1) °C. Data collected from 8–12 experiments were averaged and traces were fit to a single-exponential function. Kinetic traces were analysed using Kaleidagraph (Synergy Software).