膜
膜蛋白
外周膜蛋白
整体膜蛋白
脂质双层
囊泡相关膜蛋白8
细胞生物学
生物
拟南芥
小泡
生物物理学
生物化学
基因
突变体
作者
Martin Stuebler,Zachary A. Manzer,Han-Yuan Liu,Julia Miller,Annett Richter,S. Krishnan,Ekaterina Selivanovitch,Barituziga Banuna,Georg Jander,Erik Reimhult,Warren Zipfel,Adrienne Roeder,Miguel A. Piñeros,Susan Daniel
标识
DOI:10.1021/acsami.3c18562
摘要
The cell plasma membrane is a two-dimensional, fluid mosaic material composed of lipids and proteins that create a semipermeable barrier defining the cell from its environment. Compared with soluble proteins, the methodologies for the structural and functional characterization of membrane proteins are challenging. An emerging tool for studies of membrane proteins in mammalian systems is a "plasma membrane on a chip," also known as a supported lipid bilayer. Here, we create the "plant-membrane-on-a-chip,″ a supported bilayer made from the plant plasma membranes of Arabidopsis thaliana, Nicotiana benthamiana, or Zea mays. Membrane vesicles from protoplasts containing transgenic membrane proteins and their native lipids were incorporated into supported membranes in a defined orientation. Membrane vesicles fuse and orient systematically, where the cytoplasmic side of the membrane proteins faces the chip surface and constituents maintain mobility within the membrane plane. We use plant-membrane-on-a-chip to perform fluorescent imaging to examine protein-protein interactions and determine the protein subunit stoichiometry of FLOTILLINs. We report here that like the mammalian FLOTILLINs, FLOTILLINs expressed in Arabidopsis form a tetrameric complex in the plasma membrane. This plant-membrane-on-a-chip approach opens avenues to studies of membrane properties of plants, transport phenomena, biophysical processes, and protein-protein and protein-lipid interactions in a convenient, cell-free platform.
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