A role for amyloid in cell aggregation and biofilm formation.

A role for amyloid in cell aggregation and biofilm formation.
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DOI:
10.1371/journal.pone.0017632
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发表时间:
2011-03-08
期刊:
影响因子:
3.7
通讯作者:
Lipke PN
Lipke PN
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Garcia MC;Lee JT;Ramsook CB;Alsteens D;Dufrêne YF;Lipke PN

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酿酒酵母和白色念珠菌中的细胞粘附分子含有高度保守的淀粉样蛋白形成序列。我们现在已经使用位点特异性诱变和特定的肽干扰剂来探索白念珠菌粘附素Als5p中的淀粉样蛋白依赖性活性。淀粉样蛋白形成区域中的V326 N取代保留了二级结构和配体结合,但废除了可溶性Als5p中淀粉样蛋白原纤维的形成,并降低了细胞表面硫磺素T荧光。当在细胞表面上展示时,具有这种取代的Als5p防止粘附纳米结构域的形成以及大细胞聚集体和模型生物膜的形成。此外,淀粉样蛋白纳米结构域受外源肽的调节。淀粉样蛋白形成同源肽拯救Als5pV326N细胞的聚集和生物膜活性,并且V326N取代肽抑制Als5pWT细胞中的聚集和生物膜活性。因此,特异性位点突变、抗淀粉样蛋白抑制剂的抑制以及前淀粉样蛋白和抗淀粉样蛋白肽的序列特异性表明淀粉样蛋白形成对于纳米结构域形成和活化是必不可少的。
Cell adhesion molecules in Saccharomyces cerevisiae and Candida albicans contain amyloid-forming sequences that are highly conserved. We have now used site-specific mutagenesis and specific peptide perturbants to explore amyloid-dependent activity in the Candida albicans adhesin Als5p. A V326N substitution in the amyloid-forming region conserved secondary structure and ligand binding, but abrogated formation of amyloid fibrils in soluble Als5p and reduced cell surface thioflavin T fluorescence. When displayed on the cell surface, Als5p with this substitution prevented formation of adhesion nanodomains and formation of large cellular aggregates and model biofilms. In addition, amyloid nanodomains were regulated by exogenous peptides. An amyloid-forming homologous peptide rescued aggregation and biofilm activity of Als5pV326N cells, and V326N substitution peptide inhibited aggregation and biofilm activity in Als5pWT cells. Therefore, specific site mutation, inhibition by anti-amyloid peturbants, and sequence-specificity of pro-amyloid and anti-amyloid peptides showed that amyloid formation is essential for nanodomain formation and activation.
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