Mapping Potential Engineering Sites of Mycobacterium smegmatis porin A (MspA) to Form a Nanoreactor,ACS Sensors

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Mapping Potential Engineering Sites of Mycobacterium smegmatis porin A (MspA) to Form a Nanoreactor
ACS Sensors ( IF 8.2 ) Pub Date : 2021-06-09 , DOI: 10.1021/acssensors.1c00792
Jinyue Zhang _{1,

2} , Jiao Cao _{1,

2} , Wendong Jia _{1,

2} , Shanyu Zhang _{1,

2} , Shuanghong Yan _{1,

2} , Yuqin Wang _{1,

2} , Panke Zhang ₁ , Hong-Yuan Chen ₁ , Wenfei Li ₃ , Shuo Huang _{1,

2}

Affiliation

Protein nanopores can be engineered as nanoreactors to investigate single-molecule chemical reactions. Recent studies have demonstrated that Mycobacterium smegmatis porin A (MspA) nanopore is a superior engineering template acknowledging its geometrical advantages. However, reported engineering of MspA to form a nanoreactor has focused only on site 91 and mapping of other engineering sites have never been performed before. By taking tetrachloraurate(III) ([AuCl₄]⁻) as a model reactant, potential engineering sites within the pore constriction of MspA have been thoroughly investigated. It is discovered that the produced event amplitude is inversely correlated to the cross-sectional diameter of the pore constriction size at the engineering site, providing evidence that site 91 is actually already the optimum place to introduce the chemical reactivity. Other unavailable engineering sites, which either significantly interfere with the pore assembly or produce reactive sites facing to the pore’s exterior instead of to the pore lumen, were also spotted and discussed. All results demonstrated above have provided a complete map of engineering sites within the constriction area of MspA and may be beneficial as a reference in future engineering of corresponding nanoreactors.

中文翻译：

绘制耻垢分枝杆菌孔蛋白 A (MspA) 的潜在工程位点以形成纳米反应器

蛋白质纳米孔可以设计为纳米反应器来研究单分子化学反应。最近的研究表明，耻垢分枝杆菌孔蛋白 A (MspA) 纳米孔是一种优越的工程模板，承认其几何优势。然而，已报道的 MspA 工程形成纳米反应器仅关注站点 91，之前从未进行过其他工程站点的映射。通过服用四氯金酸盐（III）（[AuCl ₄ ] ^-) 作为模型反应物，已经彻底研究了 MspA 孔收缩内的潜在工程位点。发现产生的事件幅度与工程现场孔隙收缩尺寸的横截面直径成反比，证明现场91实际上已经是引入化学反应的最佳位置。还发现并讨论了其他不可用的工程位点，它们要么显着干扰孔组装，要么产生面向孔外部而不是孔腔的反应位点。上面展示的所有结果都提供了 MspA 收缩区域内工程位点的完整地图，可能有助于作为未来相应纳米反应器工程的参考。

更新日期：2021-06-25

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