Formation of functional, non-amyloidogenic fibres by recombinant Bacillus subtilis TasA.

Formation of functional, non-amyloidogenic fibres by recombinant Bacillus subtilis TasA.
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DOI:
10.1111/mmi.13985
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发表时间:
2018-12
影响因子:
3.6
通讯作者:
MacPhee CE
MacPhee CE
中科院分区:
生物学2区
文献类型:
--
作者:
Erskine E;Morris RJ;Schor M;Earl C;Gillespie RMC;Bromley KM;Sukhodub T;Clark L;Fyfe PK;Serpell LC;Stanley-Wall NR;MacPhee CE

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细菌生物膜是包裹在自生聚合物基质中的微生物细胞群落。在枯草芽孢杆菌生物膜基质中,TasA的细胞外纤维是必不可少的。在这里,重组表达系统允许询问TasA,揭示单体和纤维形式的TasA具有相同的二级结构,表明纤维TasA是球状单位的线性组装。重组TasA纤维自发形成,并具有从B中提取的TasA纤维的生物活性。枯草芽孢杆菌,而限制于单体形式的TasA变体是无活性的,并进行胞外蛋白水解。天然和重组TasA纤维的生物物理性质表明它们不是功能性淀粉样蛋白样纤维。由TasA纤维形成的凝胶可以在物理剪切力后恢复,这表明生物膜基质不是静态的,并且这些性质可以实现B。枯草杆菌重塑其本地环境,以响应外部线索。使用由TasA直系同源物形成的重组纤维,我们揭示了异源纤维交叉互补B的能力的物种变异性。枯草杆菌tasA缺失。这些发现表明了不同物种之间TasA纤维的生物物理要求的特异性和/或反映了生物膜基质组装所需的精确分子相互作用。
Bacterial biofilms are communities of microbial cells encased within a self‐produced polymeric matrix. In the Bacillus subtilis biofilm matrix, the extracellular fibres of TasA are essential. Here, a recombinant expression system allows interrogation of TasA, revealing that monomeric and fibre forms of TasA have identical secondary structure, suggesting that fibrous TasA is a linear assembly of globular units. Recombinant TasA fibres form spontaneously, and share the biological activity of TasA fibres extracted from B. subtilis, whereas a TasA variant restricted to a monomeric form is inactive and subjected to extracellular proteolysis. The biophysical properties of both native and recombinant TasA fibres indicate that they are not functional amyloid‐like fibres. A gel formed by TasA fibres can recover after physical shear force, suggesting that the biofilm matrix is not static and that these properties may enable B. subtilis to remodel its local environment in response to external cues. Using recombinant fibres formed by TasA orthologues we uncover species variability in the ability of heterologous fibres to cross‐complement the B. subtilis tasA deletion. These findings are indicative of specificity in the biophysical requirements of the TasA fibres across different species and/or reflect the precise molecular interactions needed for biofilm matrix assembly.
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