Laboratory of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, 37077, Göttingen, Germany.
Department of Biophysical Chemistry, Institute for Physical and Theoretical Chemistry, University of Braunschweig, 38106, Braunschweig, Germany.
Eur Biophys J. 2021 Mar;50(2):211-222. doi: 10.1007/s00249-021-01516-6. Epub 2021 Mar 31.
In the past decade, we developed various fluorescence-based methods for monitoring membrane fusion, membrane docking, distances between membranes, and membrane curvature. These tools were mainly developed using liposomes as model systems, which allows for the dissection of specific interactions mediated by, for example, fusion proteins. Here, we provide an overview of these methods, including two-photon fluorescence cross-correlation spectroscopy and intramembrane Förster energy transfer, with asymmetric labelling of inner and outer membrane leaflets and the calibrated use of transmembrane energy transfer to determine membrane distances below 10 nm. We discuss their application range and their limitations using examples from our work on protein-mediated vesicle docking and fusion.
在过去的十年中，我们开发了各种基于荧光的方法来监测膜融合、膜对接、膜间距离和膜曲率。这些工具主要是使用脂质体作为模型系统开发的，这使得可以剖析例如融合蛋白介导的特定相互作用。在这里，我们提供了这些方法的概述，包括双光子荧光交叉相关光谱法和膜内Förster 能量转移，以及对内、外膜叶的不对称标记和对跨膜能量转移的校准使用，以确定低于 10nm 的膜距离。我们使用我们在蛋白介导的囊泡对接和融合工作中的例子讨论了它们的应用范围和局限性。
