Protein folding and unfolding in microseconds to nanoseconds by experiment and simulation

The Engrailed Homeodomain protein has the highest refolding and unfolding rate constants directly observed to date. Temperature jump relaxation measurements gave a refolding rate constant of 37,500 s(-1) in water at 25 degreesC, rising to 51,000 s(-1) around 42 degreesC. The unfolding rate constant was 1,100 s(-1) in water at 25 degreesC and 205,000 s(-1) at 63 degreesC. The unfolding half-life is extrapolated to be approximate to7.5 ns at 100 degreesC, which allows real-time molecular dynamics unfolding simulations to be tested an this system at a realistic temperature. Preliminary simulations did indeed conform to unfolding on this time scale. Further, similar transition states were observed in simulations at 100 degreesC and 225 degreesC, suggesting that high-temperature simulations provide results applicable to lower temperatures.