Protein folding and unfolding in microseconds to nanoseconds by experiment and simulation
Protein folding and unfolding in microseconds to nanoseconds by experiment and simulation
复制标题
DOI：
10.1073/pnas.250473497
复制
发表时间：
2000-12-05
影响因子：
11.1
通讯作者：
Fersht, AR
中科院分区：
文献类型：
--
作者：
Mayor, U;Johnson, CM;Fersht, AR
Engrailed同源结构域蛋白具有迄今为止直接观察到的最高重折叠和解折叠速率常数。温度跃变弛豫测量给出了在25 ℃水中的重折叠速率常数为37，500 s（-1），在42 ℃左右上升到51，000 s（-1）。在25 ℃和63 ℃下，在水中的解折叠速率常数分别为1，100 s（-1）和205，000 s（-1）。解折叠半衰期外推到约为7.5 ns在100 degreesC，这使得实时分子动力学解折叠模拟测试在这个系统在一个现实的温度。初步的模拟确实符合这个时间尺度上的展开。此外，在100摄氏度和225摄氏度的模拟中观察到类似的过渡态，这表明高温模拟提供了适用于较低温度的结果。
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.