Institute of Physical Chemistry, Georg-August-University, Tammannstrasse 6, 37077 Göttingen, Germany.
J Am Chem Soc. 2013 Aug 21;135(33):12176-9. doi: 10.1021/ja404071z. Epub 2013 Aug 9.
A major goal in neurophysiology and research on enveloped viruses is to understand and control the biology and physics of membrane fusion and its inhibition as a function of lipid and protein composition. This poses an experimental challenge in the realization of fast and reliable assays that allow us, with a minimal use of fluorescent or radioactive labels, to identify the different stages of membrane-membrane interaction ranging from docking to complete membrane merging. Here, an optical two-dimensional fusion assay based on monodisperse membrane-coated microspheres is introduced, allowing unequivocal assignment of docking and membrane fusion. The hard-sphere fluid captures and quantifies relevant stages of membrane fusion and its inhibition without interference from aggregation, liposome rupture, extensive fluorescence labeling, and light scattering. The feasibility of the approach is demonstrated by using an established model system based on coiled-coil heterodimers formed between two opposing membrane-coated microspheres.
神经生理学和包膜病毒研究的主要目标是理解和控制膜融合及其抑制的生物学和物理学特性，作为脂质和蛋白质组成的函数。这在实现快速可靠的测定方面提出了一个实验挑战，使我们能够在最小使用荧光或放射性标记的情况下，确定从对接至完全膜融合的膜-膜相互作用的不同阶段。本文介绍了一种基于单分散膜包被微球的光学二维融合测定法，允许明确分配对接和膜融合。硬球流体捕获和量化了膜融合及其抑制的相关阶段，而不会受到聚集、脂质体破裂、广泛荧光标记和光散射的干扰。通过使用基于两个相反的膜包被微球之间形成的卷曲螺旋异二聚体的既定模型系统，证明了该方法的可行性。
