小柯机器人

美国东北大学Meni Wanunu课题组取得一项新突破。他们的研究实现利用纳米孔对全长蛋白质进行单向单个传输。该项研究成果发表在2023年1月9日出版的《自然—生物技术》上。

研究人员研发了一种无需酶的缓慢、单向运输全长蛋白质通过纳米孔的方法。研究表明耐化学性的生物纳米孔，α溶血素（最窄部分直径为1.4nm）和高浓度氯化胍缓冲液的组合能够实现由电渗效应推动的单向单个蛋白质转运。研究表明，平均蛋白质易位速度取决于施加的电压，易位时间取决于蛋白长度，类似于ssDNA易位动力学。使用监督式机器学习分类器，研究人员发现单易位事件包含足够的信息来区分其线程方向和精度大于90的标识。将基因编码的带电肽尾或DNA标签添加到蛋白质中时，蛋白质的捕获率显著增加。

据介绍，肽或蛋白质穿过纳米孔时的电流阻断可在生物传感器应用中作为分子指纹图谱。然而，全长蛋白质穿过纳米孔仅通过酶促反应解折叠和易位来实现。

附：英文原文

Title: Unidirectional single-file transport of full-length proteins through a nanopore

Author: Yu, Luning, Kang, Xinqi, Li, Fanjun, Mehrafrooz, Behzad, Makhamreh, Amr, Fallahi, Ali, Foster, Joshua C., Aksimentiev, Aleksei, Chen, Min, Wanunu, Meni

Issue&Volume: 2023-01-09

Abstract: The electrical current blockade of a peptide or protein threading through a nanopore can be used as a fingerprint of the molecule in biosensor applications. However, threading of full-length proteins has only been achieved using enzymatic unfolding and translocation. Here we describe an enzyme-free approach for unidirectional, slow transport of full-length proteins through nanopores. We show that the combination of a chemically resistant biological nanopore, α-hemolysin (narrowest part is ~1.4nm in diameter), and a high concentration guanidinium chloride buffer enables unidirectional, single-file protein transport propelled by an electroosmotic effect. We show that the mean protein translocation velocity depends linearly on the applied voltage and translocation times depend linearly on length, resembling the translocation dynamics of ssDNA. Using a supervised machine-learning classifier, we demonstrate that single-translocation events contain sufficient information to distinguish their threading orientation and identity with accuracies larger than 90%. Capture rates of protein are increased substantially when either a genetically encoded charged peptide tail or a DNA tag is added to a protein.

DOI: 10.1038/s41587-022-01598-3

Source: https://www.nature.com/articles/s41587-022-01598-3

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
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