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首页> 外文期刊>Cellular Physiology and Biochemistry >Plasma membrane plasticity of Xenopus laevis oocyte imaged with atomic force microscopy
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Plasma membrane plasticity of Xenopus laevis oocyte imaged with atomic force microscopy

机译：原子力显微镜成像的非洲爪蟾卵母细胞质膜可塑性

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Proteins are known to form functional clusters in plasma membranes. In order to identify individual proteins within clusters we developed a method to visualize by atomic force microscopy (AFM) the cytoplasmic surface of native plasma membrane, excised from Xenopus laevis oocyte and spread on poly-L-lysine coated glass. After removal of the vitelline membrane intact oocytes were brought in contact with coated glass and then rolled off. Inside-out oriented plasma membrane patches left at the glass surface were first identified with the lipid fluorescent marker FM1-43 and then scanned by AFM. Membrane patches exhibiting the typical phospholipid bilayer height of 5 nm showed multiple proteins, protruding from the inner surface of the membrane, with heights of 5 to 20 nm. Modelling plasma membrane proteins as spherical structures embedded in the lipid bilayer and protruding into the cytoplasm allowed an estimation of the respective molecular masses. Proteins ranged from 35 to 2,000 kDa with a peak value of 280 kDa. The most frequently found membrane protein structure (40/ mu m(2)) had a total height of 10 nm and an estimated molecular mass of 280 kDa. Membrane proteins were found firmly attached to the poly-L-lysine coated glass surface while the lipid bilayer was found highly mobile. We detected protein structures with distinguishable subunits of still unknown identity. Since X. laevis oocyte is a generally accepted expression system for foreign proteins, this method could turn out to be useful to structurally identify specific proteins in their native environment at the molecular level. Copyright (C) 2000 S. Karger AG, Basel. [References: 18]
机译：已知蛋白质会在质膜上形成功能簇。为了鉴定簇中的单个蛋白质，我们开发了一种方法，通过原子力显微镜（AFM）可视化天然细胞膜的细胞质表面，该表面从非洲爪蟾卵母细胞中切下，并铺在聚L-赖氨酸涂层的玻璃上。除去卵黄膜后，使完整的卵母细胞与涂覆的玻璃接触，然后滚下来。首先用脂质荧光标记FM1-43识别留在玻璃表面的由内而外定向的质膜斑块，然后通过AFM进行扫描。表现出典型的磷脂双层高度为5 nm的膜片显示出多种蛋白，从膜的内表面突出，高度为5至20 nm。将质膜蛋白建模为嵌入脂质双分子层并伸入细胞质的球形结构，可以估算各自的分子质量。蛋白质范围为35至2,000 kDa，峰值为280 kDa。最常见的膜蛋白结构（40 /μm（2））总高度为10 nm，估计分子量为280 kDa。发现膜蛋白牢固地附着在聚-L-赖氨酸包被的玻璃表面上，而脂质双层被发现具有高度的流动性。我们检测到具有未知身份的可区分亚基的蛋白质结构。由于X. laevis卵母细胞是外来蛋白质的普遍接受的表达系统，因此该方法可用于在分子水平上在其天然环境中结构鉴定特定蛋白质。版权所有（C）2000 S.Karger AG，巴塞尔。 [参考：18]

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