Congo Red Interactions with Curli-Producing E. coli and Native Curli Amyloid Fibers.
Congo Red Interactions with Curli-Producing E. coli and Native Curli Amyloid Fibers.
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DOI：
10.1371/journal.pone.0140388
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Cegelski L
中科院分区：
文献类型：
--
作者：
Reichhardt C;Jacobson AN;Maher MC;Uang J;McCrate OA;Eckart M;Cegelski L
微生物产生功能性淀粉样蛋白，可以在体内和体外进行检查和操作。大肠杆菌组装称为 Curli 的细胞外粘附淀粉样纤维，介导粘附并促进生物膜形成。我们表征了标志性淀粉样蛋白染料刚果红与卷曲大肠杆菌和分离的卷曲纤维的染料结合特性。刚果红与卷曲的整个细胞结合，不抑制生长，并且可用于相对量化全细胞的卷曲。使用表面等离子共振，我们测量了刚果红与 curli 的结合和解离动力学。此外，我们确定刚果红与 Curli 的结合是 pH 依赖性的，并且 CsgA 蛋白中的组氨酸残基不会影响刚果红结合。我们对大肠杆菌菌株 MC4100（大肠杆菌淀粉样蛋白生物发生研究最常用的菌株）的研究结果为比较刚果红结合对其他大肠杆菌菌株和淀粉样蛋白产生生物体的影响提供了一个起点。
Microorganisms produce functional amyloids that can be examined and manipulated in vivo and in vitro. Escherichia coli assemble extracellular adhesive amyloid fibers termed curli that mediate adhesion and promote biofilm formation. We have characterized the dye binding properties of the hallmark amyloid dye, Congo red, with curliated E. coli and with isolated curli fibers. Congo red binds to curliated whole cells, does not inhibit growth, and can be used to comparatively quantify whole-cell curliation. Using Surface Plasmon Resonance, we measured the binding and dissociation kinetics of Congo red to curli. Furthermore, we determined that the binding of Congo red to curli is pH-dependent and that histidine residues in the CsgA protein do not influence Congo red binding. Our results on E. coli strain MC4100, the most commonly employed strain for studies of E. coli amyloid biogenesis, provide a starting point from which to compare the influence of Congo red binding in other E. coli strains and amyloid-producing organisms.