Khadria Ambalika S, Senes Alessandro
Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706.
Biopolymers. 2015 Jul;104(4):247-64. doi: 10.1002/bip.22667.
Förster resonance energy transfer (FRET) has been widely used as a spectroscopic tool in vitro to study the interactions between transmembrane (TM) helices in detergent and lipid environments. This technique has been instrumental to many studies that have greatly contributed to quantitative understanding of the physical principles that govern helix-helix interactions in the membrane. These studies have also improved our understanding of the biological role of oligomerization in membrane proteins. In this review, we focus on the combinations of fluorophores used, the membrane mimetic environments, and measurement techniques that have been applied to study model systems as well as biological oligomeric complexes in vitro. We highlight the different formalisms used to calculate FRET efficiency and the challenges associated with accurate quantification. The goal is to provide the reader with a comparative summary of the relevant literature for planning and designing FRET experiments aimed at measuring TM helix-helix associations.
荧光共振能量转移（FRET）已被广泛用作体外光谱工具，用于研究去污剂和脂质环境中跨膜（TM）螺旋之间的相互作用。这项技术对许多研究起到了重要作用，这些研究极大地促进了对控制膜中螺旋 - 螺旋相互作用的物理原理的定量理解。这些研究还增进了我们对膜蛋白中寡聚化生物学作用的理解。在这篇综述中，我们重点关注用于研究体外模型系统以及生物寡聚复合物的荧光团组合、膜模拟环境和测量技术。我们强调了用于计算FRET效率的不同形式主义以及与准确定量相关的挑战。目的是为读者提供相关文献的比较性总结，以便规划和设计旨在测量TM螺旋 - 螺旋关联的FRET实验。
