Schuler Benjamin
Department of Biochemistry, University of Zürich, Zürich, Switzerland.
Methods Mol Biol. 2007;350:115-38. doi: 10.1385/1-59745-189-4:115.
Protein folding is a process characterized by a large degree of conformational heterogeneity. In such cases, classical experimental methods yield only mean values, averaged over large ensembles of molecules. The microscopic distributions of conformations, trajectories, or sequences of events often remain unknown, and with them the underlying molecular mechanisms. Signal averaging can be avoided by observing individual molecules. A particularly versatile method is highly sensitive fluorescence detection. In combination with Förster resonance energy transfer, distances and conformational dynamics can be investigated in single molecules. This chapter introduces the practical aspects of applying this method to protein folding.
蛋白质折叠是一个具有高度构象异质性的过程。在这种情况下，经典实验方法只能得出在大量分子集合上平均得到的平均值。构象、轨迹或事件序列的微观分布通常仍然未知，与之相关的潜在分子机制也未知。通过观察单个分子可以避免信号平均。一种特别通用的方法是高灵敏度荧光检测。结合Förster共振能量转移，可以在单分子中研究距离和构象动力学。本章介绍了将该方法应用于蛋白质折叠的实际操作要点。
