Plant Membrane-On-A-Chip: A Platform for Studying Plant Membrane Proteins and Lipids

被引:0
作者
Stuebler, Martin [1 ,2 ]
Manzer, Zachary A. [1 ]
Liu, Han-Yuan [1 ]
Miller, Julia [3 ]
Richter, Annett [4 ]
Krishnan, Srinivasan [4 ]
Selivanovitch, Ekaterina [1 ]
Banuna, Barituziga [1 ]
Jander, Georg [4 ]
Reimhult, Erik [2 ]
Zipfel, Warren R. [5 ]
Roeder, Adrienne H. K. [3 ,6 ]
Pineros, Miguel A. [3 ,7 ]
Daniel, Susan [1 ]
机构
[1] Cornell Univ, RF Smith Sch Chem & Biomol Engn, Ithaca, NY 14853 USA
[2] Univ Nat Resources & Life Sci, A-1180 Vienna, Austria
[3] Cornell Univ, Sch Integrat Plant Sci, Ithaca, NY 14853 USA
[4] Boyce Thompson Inst Plant Res, Ithaca, NY 14853 USA
[5] Cornell Univ, Meinig Sch Biomed Engn, Ithaca, NY 14853 USA
[6] Cornell Univ, Weill Inst Cell & Mol Biol, Ithaca, NY 14853 USA
[7] ARS USDA, Robert W Holley Ctr Agr & Hlth, Ithaca, NY 14853 USA
基金
美国国家科学基金会;
关键词
protoplast; plant cell membrane; supportedlipid bilayer; bioelectronics; plant membrane; PLASMA-MEMBRANE; LATERAL DIFFUSION; BROWNIAN-MOTION; ARABIDOPSIS; BILAYERS; TRACKING; MOBILITY; MICRODOMAINS; PROTOPLASTS; EXPRESSION;
D O I
10.1021/acsami.3c18562
中图分类号
TB3 [工程材料学];
学科分类号
0805 ; 080502 ;
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.
引用
收藏
页码:20092 / 20104
页数:13
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