Dynamics of long hyaluronic acid chains through conical nanochannels for characterizing enzyme reactions in confined spaces,Nanoscale

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Dynamics of long hyaluronic acid chains through conical nanochannels for characterizing enzyme reactions in confined spaces
Nanoscale ( IF 5.8 ) Pub Date : 2020/02/24 , DOI: 10.1039/d0nr00645a
Tianji Ma _{1,

2,

3,

4} , Jean-Marc Janot _{1,

2,

3,

4} , Sébastien Balme _{1,

2,

3,

4}

Affiliation

This research reports the transport behaviors of long flexible polymers (hyaluronic acid) through long conical track-etched nanochannels with and without grafted enzymes. The impacts of the channel diameter and the polymer regimes in solution (dilute and semi-dilute) have been investigated. Without enzymes, the experimental results can be well explained by the analytical models of the scaling law of de Gennes. Then, the corresponding enzymes (hyaluronidase) were grafted inside the channel. When enzymes are located at the base side, polymers get degraded at the entrance and the degraded products are detected. When enzymes are grafted at the tip side, the extension of translocation duration due to the binding of substrate–enzyme is observed. This is for the first time that the enzymatic degradation reactions are characterized in situ at the single molecule level by nanopore technology.

中文翻译：

通过圆锥形纳米通道的透明质酸长链动力学，用于表征密闭空间中的酶反应

这项研究报告了长柔性聚合物（透明质酸）通过带有或不带有接枝酶的长圆锥形轨迹蚀刻纳米通道的传输行为。研究了通道直径和溶液（稀溶液和半稀溶液）中聚合物形式的影响。没有酶，de Gennes比例定律的分析模型可以很好地解释实验结果。然后，将相应的酶（透明质酸酶）移植到通道内。当酶位于碱基侧时，聚合物在入口处降解，并检测到降解产物。当酶在末端接枝时，观察到由于底物-酶结合而导致的转运时间延长。这是首次表征酶促降解反应通过纳米孔技术在单分子水平进行原位检测。

更新日期：2020-04-03

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