Niels Bohr Institute, University of Copenhagen, Blegdamsvej 17, 2100 København Ø, Denmark.
Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Potsdam, Germany.
Nano Lett. 2023 Apr 26;23(8):3377-3384. doi: 10.1021/acs.nanolett.3c00371. Epub 2023 Apr 11.
Many cellular processes involve the lateral organization of integral and peripheral membrane proteins into nanoscale domains. Despite the biological significance, the mechanisms that facilitate membrane protein clustering into nanoscale lipid domains remain enigmatic. In cells, the analysis of membrane protein phase affinity is complicated by the size and temporal nature of ordered and disordered lipid domains. To overcome these limitations, we developed a method for delivering membrane proteins from transfected cells into phase-separated model membranes that combines optical trapping with thermoplasmonic-mediated membrane fusion and confocal imaging. Using this approach, we observed clear phase partitioning into the liquid disordered phase following the transfer of GFP-tagged influenza hemagglutinin and neuraminidase from transfected cell membranes to giant unilamellar vesicles. The generic platform presented here allows investigation of the phase affinity of any plasma membrane protein which can be labeled or tagged with a fluorescent marker.
许多细胞过程涉及将整合膜蛋白和外周膜蛋白侧向组织成纳米尺度的域。尽管具有生物学意义，但促进膜蛋白聚类成纳米尺度脂质域的机制仍然是个谜。在细胞中，由于有序和无序脂质域的大小和时间性质，膜蛋白相亲和力的分析变得复杂。为了克服这些限制，我们开发了一种将转染细胞中的膜蛋白递送到相分离模型膜中的方法，该方法结合了光镊与热等离子体介导的膜融合和共焦成像。使用这种方法，我们观察到 GFP 标记的流感血凝素和神经氨酸酶从转染细胞膜转移到巨大的单层囊泡后，明显地分配到液体无序相中。这里提出的通用平台允许研究任何可以用荧光标记物标记或标记的质膜蛋白的相亲和力。
