中国科学院化学研究所吴海臣和刘蕾共同合作，近期取得重要工作进展。他们研究开发了基于主客体相互作用辅助纳米孔传感的肽测序技术。相关研究成果2023年11月13日在线发表于《自然—方法学》杂志上。

据介绍，目前仍然需要具有改进的灵敏度和高通量的直接蛋白质测序技术。

研究人员提出了一种基于酶切和主-客体相互作用辅助纳米孔传感的肽测序替代方法。研究人员发现，在含有苯丙氨酸的肽的特定位置上，任何蛋白原氨基酸的身份都可以通过在瓜血凝素存在的情况下通过α-溶血素纳米孔转运时的电流阻塞来确定。在此基础上，研究人员进一步提出了肽测序的概念验证，方法是用羧肽酶从肽的C末端依次切割氨基酸，然后用纳米孔中的肽探针确定它们的身份和序列。

总之，随着未来的优化，这一研究结果指向了一种不同的基于纳米孔的蛋白质测序方法。

附：英文原文

Title: Peptide sequencing based on host–guest interaction-assisted nanopore sensing

Author: Zhang, Yun, Yi, Yakun, Li, Ziyi, Zhou, Ke, Liu, Lei, Wu, Hai-Chen

Issue&Volume: 2023-11-13

Abstract: Direct protein sequencing technologies with improved sensitivity and throughput are still needed. Here, we propose an alternative method for peptide sequencing based on enzymatic cleavage and host–guest interaction-assisted nanopore sensing. We serendipitously discovered that the identity of any proteinogenic amino acid in a particular position of a phenylalanine-containing peptide could be determined via current blockage during translocation of the peptide through α-hemolysin nanopores in the presence of cucurbit[7]uril. Building upon this, we further present a proof-of-concept demonstration of peptide sequencing by sequentially cleaving off amino acids from C terminus of a peptide with carboxypeptidases, and then determining their identities and sequence with a peptide probe in nanopore. With future optimization, our results point to a different way of nanopore-based protein sequencing.

DOI: 10.1038/s41592-023-02095-4

Source: https://www.nature.com/articles/s41592-023-02095-4