Silvius John R, Nabi Ivan Robert
Department of Biochemistry, McGill University, Montréal, Québec, Canada.
Mol Membr Biol. 2006 Jan-Feb;23(1):5-16. doi: 10.1080/09687860500473002.
Measurements of contact-dependent fluorescence quenching and of fluorescence resonance energy transfer (FRET) within bilayers provide information concerning the spatial relationships between molecules on distance scales of a few nm or up a few tens of nm, respectively, and are therefore well suited to detect the presence and composition of membrane microdomains. As described in this review, techniques based on fluorescence quenching and FRET have been used to demonstrate the formation of nanoscale liquid-ordered domains in cholesterol-containing model membranes under physiological conditions, and to investigate the structural features of lipids and proteins that influence their partitioning between liquid-ordered and liquid-disordered domains. FRET-based methods have also been used to test for the presence of 'raft' microdomains in the plasma membranes of mammalian cells. We discuss the sometimes divergent findings of these studies, possible modifications to the 'raft hypothesis' suggested by studies using FRET and other techniques, and the further potential of FRET-based methods to test and to refine current models of the nature and organization of membrane microdomains.
双层膜内接触依赖性荧光猝灭和荧光共振能量转移（FRET）的测量分别提供了关于分子在几纳米到几十纳米距离尺度上的空间关系的信息，因此非常适合用于检测膜微区的存在和组成。如本综述所述，基于荧光猝灭和FRET的技术已被用于证明在生理条件下含胆固醇的模型膜中纳米级液相有序域的形成，并研究影响脂质和蛋白质在液相有序域和液相无序域之间分配的结构特征。基于FRET的方法也被用于检测哺乳动物细胞质膜中“筏”微区的存在。我们讨论了这些研究中有时存在分歧的结果、使用FRET和其他技术的研究所提出的对“筏假说”的可能修正，以及基于FRET的方法在测试和完善当前膜微区性质和组织模型方面的进一步潜力。
