Forces in yeast flocculation.
Forces in yeast flocculation.
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DOI：
10.1039/c4nr06315e
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发表时间：
2015-02-07
期刊：
影响因子：
6.7
通讯作者：
Dufrêne YF
中科院分区：
文献类型：
--
作者：
El-Kirat-Chatel S;Beaussart A;Vincent SP;Abellán Flos M;Hols P;Lipke PN;Dufrêne YF
在面包酵母酿酒酵母中，细胞与细胞的粘附（“絮凝”）是由称为絮凝蛋白（Flo）蛋白的凝集素样蛋白家族赋予的。了解絮凝蛋白的粘附和机械特性对于理解酵母粘附机制很重要，并且可能有助于控制生物技术中的酵母行为。我们使用单分子和单细胞原子力显微镜 (AFM) 来探索参与酵母絮凝的纳米级力，重点关注 Flo1 作为絮凝素原型的作用。使用甘露糖标记的 AFM 吸头，我们在表达 Flo1 的细胞上检测到单个絮凝蛋白，表明它们广泛暴露在细胞表面。当受到力时，各个 Flo1 蛋白表现出两种不同的力响应，即弱的凝集素结合力和强的解折叠力，反映了力诱导的疏水性串联重复序列的延伸。我们证明细胞-细胞粘附键还涉及多个弱凝集素相互作用和强展开力，两者都与 Flo1 分子相关。单分子和单细胞数据与微尺度细胞粘附行为相关，强烈表明 Flo1 力学对于酵母絮凝至关重要。这些结果支持这样一个模型，在该模型中，酵母絮凝中不仅涉及弱的凝集素-糖相互作用，而且还涉及蛋白质解折叠产生的强疏水相互作用。
In the baker's yeast Saccharomyces cerevisiae, cell-cell adhesion (“flocculation”) is conferred by a family of lectin-like proteins known as the flocculin (Flo) proteins. Knowledge of the adhesive and mechanical properties of flocculins is important for understanding the mechanisms of yeast adhesion, and may help controlling yeast behaviour in biotechnology. We use single-molecule and single-cell atomic force microscopy (AFM) to explore the nanoscale forces engaged in yeast flocculation, focusing on the role of Flo1 as a prototype of flocculins. Using AFM tips labelled with mannose, we detect single flocculins on Flo1-expressing cells, showing they are widely exposed on the cell surface. When subjected to force, individual Flo1 proteins display two distinct force responses, i.e. weak lectin binding forces and strong unfolding forces reflecting the force-induced extension of hydrophobic tandem repeats. We demonstrate that cell-cell adhesion bonds also involve multiple weak lectin interactions together with strong unfolding forces, both associated with Flo1 molecules. Single-molecule and single-cell data correlate with microscale cell adhesion behaviour, suggesting strongly that Flo1 mechanics is critical for yeast flocculation. These results favour a model in which not only weak lectin-sugar interactions are involved in yeast flocculation but also strong hydrophobic interactions resulting from protein unfolding.
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