Membrane fusion mediated by peptidic SNARE protein analogues: Evaluation of FRET-based bulk leaflet mixing assays,Journal of Peptide Science

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Membrane fusion mediated by peptidic SNARE protein analogues: Evaluation of FRET-based bulk leaflet mixing assays
Journal of Peptide Science ( IF 1.8 ) Pub Date : 2021-04-06 , DOI: 10.1002/psc.3327
Barbara E Hubrich ₁ , Jan-Dirk Wehland ₁ , Mike C Groth ₁ , Anastasiya Schirmacher ₁ , Raphael Hubrich ₁ , Claudia Steinem ₁ , Ulf Diederichsen ₁

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Peptide-mediated membrane fusion is frequently studied with in vitro bulk leaflet mixing assays based on Förster resonance energy transfer (FRET). In these, customized liposomes with fusogenic peptides are equipped with lipids which are labeled with fluorophores that form a FRET pair. Since FRET is dependent on distance and membrane fusion comes along with lipid mixing, the assays allow for conclusions on the membrane fusion process. The experimental outcome of these assays, however, greatly depends on the applied parameters. In the present study, the influence of the peptides, the size of liposomes, their lipid composition and the liposome stoichiometry on the fusogenicity of liposomes are evaluated. As fusogenic peptides, soluble N-ethylmaleimide-sensitive-factor attachment receptor (SNARE) protein analogues featuring artificial recognition units attached to the native SNARE transmembrane domains are used. The work shows that it is important to control these parameters in order to be able to properly investigate the fusion process and to prevent undesired effects of aggregation.

中文翻译：

由肽 SNARE 蛋白类似物介导的膜融合：基于 FRET 的散装传单混合测定的评估

肽介导的膜融合经常使用基于 Förster 共振能量转移 (FRET) 的体外散装传单混合测定进行研究。在这些中，带有融合肽的定制脂质体配备有脂质，这些脂质用形成 FRET 对的荧光团标记。由于 FRET 取决于距离，并且膜融合伴随着脂质混合，因此这些分析可以得出关于膜融合过程的结论。然而，这些测定的实验结果在很大程度上取决于应用的参数。在本研究中，评估了肽、脂质体的大小、脂质组成和脂质体化学计量对脂质体融合性的影响。作为融合肽，可溶性N-乙基马来酰亚胺敏感因子附着受体 (SNARE) 蛋白质类似物的特征是人工识别单元与天然 SNARE 跨膜结构域相连。工作表明，重要的是控制这些参数，以便能够正确研究融合过程并防止聚集的不良影响。

更新日期：2021-06-04

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