Quantification of membrane geometry and protein sorting on cell membrane protrusions using fluorescence microscopy.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-04-16 DOI:10.1016/bs.mie.2024.01.023
Shilong Yang, Zheng Shi
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Abstract

Plasma membranes are flexible and can exhibit numerous shapes below the optical diffraction limit. The shape of cell periphery can either induce or be a product of local protein density changes, encoding numerous cellular functions. However, quantifying membrane curvature and the ensuing sorting of proteins in live cells remains technically demanding. Here, we demonstrate the use of simple widefield fluorescence microscopy to study the geometrical properties (i.e., radius, length, and number) of thin membrane protrusions. Importantly, the quantification of protrusion radius establishes a platform for studying the curvature preferences of membrane proteins.

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利用荧光显微镜对细胞膜突起的膜几何形状和蛋白质分类进行量化。
质膜具有柔韧性,可呈现出低于光学衍射极限的多种形状。细胞外围的形状可以诱导局部蛋白质密度的变化,也可以是这种变化的产物,从而编码多种细胞功能。然而,量化活细胞中的膜曲率和随之而来的蛋白质分拣仍然是一项技术要求很高的工作。在这里,我们展示了使用简单的宽场荧光显微镜来研究薄膜突起的几何特性(即半径、长度和数量)。重要的是,突起半径的量化为研究膜蛋白的曲率偏好建立了一个平台。
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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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