Unité de Chimie des Interfaces, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.
Nat Chem Biol. 2009 Nov;5(11):857-62. doi: 10.1038/nchembio.220. Epub 2009 Sep 20.
Here we report on in vivo measurement of the mechanical behavior of a cell surface sensor using single-molecule atomic force microscopy. We focus on the yeast wall stress component sensor Wsc1, a plasma membrane protein that is thought to function as a rigid probe of the cell wall status. We first map the distribution of individual histidine-tagged sensors on living yeast cells by scanning the cell surface with atomic force microscopy tips carrying nitrilotriacetate groups. We then show that Wsc1 behaves like a linear nanospring that is capable of resisting high mechanical force and of responding to cell surface stress. Both a genomic pmt4 deletion and the insertion of a stretch of glycines in Wsc1 result in substantial alterations in protein spring properties, supporting the important role of glycosylation at the extracellular serine/threonine-rich region.
在这里，我们报告了使用单分子原子力显微镜对细胞表面传感器的机械行为进行体内测量。我们专注于酵母细胞壁应力分量传感器 Wsc1，这是一种被认为作为细胞壁状态刚性探针发挥功能的质膜蛋白。我们首先通过用携带氮三乙酸基团的原子力显微镜针尖扫描细胞表面，绘制活酵母细胞上单个组氨酸标记传感器的分布图谱。然后，我们表明 Wsc1 表现为一种线性纳米弹簧，能够抵抗高机械力并响应细胞表面应力。基因组 pmt4 缺失和 Wsc1 中插入一段甘氨酸都会导致蛋白质弹簧特性的实质性改变，这支持了在细胞外丝氨酸/苏氨酸丰富区域进行糖基化的重要作用。
