SUBMILLISECOND EVENTS IN PROTEIN-FOLDING
SUBMILLISECOND EVENTS IN PROTEIN-FOLDING
复制标题
DOI：
10.1073/pnas.92.23.10668
复制
发表时间：
1995-11-07
影响因子：
11.1
通讯作者：
FERSHT, AR
中科院分区：
文献类型：
--
作者：
NOLTING, B;GOLBIK, R;FERSHT, AR
蛋白质折叠的途径，现在正在分析的分辨率个别残基的动力学测量适当的工程突变体。通常用于研究折叠的动力学方法通常限于大于或等于1 ms的时间范围，因为变性蛋白质的折叠通常通过将它们与有利于折叠的缓冲液混合来启动，并且快速混合实验的死时间约为1毫秒。我们现在表明，蛋白质折叠的研究可以扩展到微秒的时间区域，通过使用一个合适的蛋白质的冷展开状态的温度跳跃测量。我们能够检测芽孢杆菌RNA酶的多肽抑制剂芽孢杆菌RNA酶抑制剂突变体折叠的早期事件。从光谱和Phi值分析的快速相的初步表征表明，它是一个过渡之间的两个相对溶剂暴露状态与小的结构合并。
The pathway of protein folding is now being analyzed at the resolution of individual residues by kinetic measurements on suitably engineered mutants. The kinetic methods generally employed for studying folding are typically limited to the time range of greater than or equal to 1 ms because the folding of denatured proteins is usually initiated by mixing them with buffers that favor folding, and the dead time of rapid mixing experiments is about a millisecond. We now show that the study of protein folding may be extended to the microsecond time region by using temperature-jump measurements on the cold-unfolded state of a suitable protein. We are able to detect early events in the folding of mutants of barstar, the polypeptide inhibitor of barnase. A preliminary characterization of the fast phase from spectroscopic and Phi-value analysis indicates that it is a transition between two relatively solvent-exposed states with little consolidation of structure.