Castorph S, Schwarz Henriques S, Holt M, Riedel D, Jahn R, Salditt T
Institut für Röntgenphysik, Georg-August-Universität Göttingen, Germany.
Eur Phys J E Soft Matter. 2011 Jun;34(6):63. doi: 10.1140/epje/i2011-11063-2. Epub 2011 Jun 27.
The size polydispersity distribution of synaptic vesicles (SVs) is characterized under quasi-physiological conditions by dynamic light scattering (DLS). Highly purified fractions of SVs obtained from rat brain still contain a small amount of larger contaminant structures, which can be quantified by DLS and further reduced by asymmetric-flow field-flow (AFFF) fractionation. The intensity autocorrelation functions g (2)(τ) recorded from these samples are analyzed by a constrained regularization method as well as by an alternative direct modeling approach. The results are in quantitative agreement with the polydispersity obtained from cryogenic electron microscopy of vitrified SVs. Next, different vesicle fusion assays based on samples composed of SVs and small unilamellar proteoliposomes with the fusion proteins syntaxin 1 and SNAP-25A are characterized by DLS. The size increase of the proteoliposomes due to SNARE-dependent fusion with SVs is quantified by DLS under quasi-physiological conditions.
在准生理条件下,通过动态光散射(DLS)对突触小泡(SVs)的大小多分散性分布进行表征。从大鼠脑获得的高度纯化的SVs组分仍含有少量较大的污染物结构,这些结构可通过DLS进行定量,并通过不对称流场流(AFFF)分级进一步减少。对这些样品记录的强度自相关函数g(2)(τ)采用约束正则化方法以及另一种直接建模方法进行分析。结果与通过玻璃化SVs的低温电子显微镜获得的多分散性在数量上一致。接下来,基于由SVs和带有融合蛋白 syntaxin 1和SNAP-25A的小单层蛋白脂质体组成的样品的不同囊泡融合测定通过DLS进行表征。在准生理条件下,通过DLS对由于SNARE依赖的与SVs融合导致的蛋白脂质体大小增加进行定量。