Kale Justin, Chi Xiaoke, Leber Brian, Andrews David
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Canada; Department of Biological Sciences, Sunnybrook Research Institute, Toronto, Canada.
Department of Biological Sciences, Sunnybrook Research Institute, Toronto, Canada; Department of Chemical Biology, McMaster University, Hamilton, Canada.
Methods Enzymol. 2014;544:1-23. doi: 10.1016/B978-0-12-417158-9.00001-7.
The Bcl-2 family proteins control apoptosis by regulation of outer mitochondrial membrane permeabilization. Studying the Bcl-2 family is particularly difficult because the functional interactions that regulate apoptosis occur at or within intracellular membranes. Compared to other biophysical methods, fluorescence spectroscopy is well suited to study membrane-bound proteins as experiments can be performed with intact membranes and at protein concentrations similar to those found in cells. For these reasons, fluorescence spectroscopy has been particularly useful in studying the regulation of membrane permeabilization by Bcl-2 family proteins. Here, we discuss four fluorescence-based assays used to study protein dynamics at membranes, with a focus on how these techniques can be used to study the Bcl-2 family proteins.
Bcl-2家族蛋白通过调节线粒体外膜通透性来控制细胞凋亡。研究Bcl-2家族特别困难，因为调节细胞凋亡的功能相互作用发生在细胞内膜上或内膜内。与其他生物物理方法相比，荧光光谱非常适合研究膜结合蛋白，因为实验可以在完整膜上进行，并且蛋白质浓度与细胞内的浓度相似。由于这些原因，荧光光谱在研究Bcl-2家族蛋白对膜通透性的调节方面特别有用。在这里，我们讨论四种基于荧光的测定方法，用于研究膜上的蛋白质动力学，重点是这些技术如何用于研究Bcl-2家族蛋白。
