南京大学黄硕团队宣布他们研究出固定在纳米孔上的逐步缩短的肽的顺序读取。相关论文于2026年7月29日发表于国际顶尖学术期刊《自然》杂志上。

在这里，课题组人员介绍了一种基于纳米孔的策略，称为瞬态孔分析物环（tPAL）。小组开发了一种工程耻垢分枝杆菌孔蛋白A （MspA）纳米孔，该纳米孔由镍离子结合的硝基三乙酸（NTA-Ni）适配器和靶肽双重修饰。这种独特的传感配置可以精确识别N通过多次重读来终止固定化肽。在胆固醇化氨基肽酶的帮助下，固定化肽可以按单氨基酸增量顺序缩短，产生序列依赖的、逐步的、窄分布的信号改变，为肽序列解码提供线索。他们的策略实现了单氨基酸分辨率，并有效识别单氨基酸突变，翻译后修饰和非天然氨基酸插入-表明其在纳米孔蛋白质组学和手性肽分析中的通用性。

据了解，肽序列的准确解码在蛋白质组学中至关重要。然而，由于肽的组成和结构的复杂性，实现这一目标是一项技术挑战。受纳米孔核酸测序成功启发的研究表明，基于纳米孔的技术也可以应用于肽测序。关键是在顺序纳米孔读出过程中产生窄分布、一致和序列相关的事件。

附：英文原文

Title: Sequential reading of a stepwise-shortened peptide immobilized on nanopore

Author: Chen, Jialu, Zhang, Hanhan, Wang, Kefan, Li, Tian, Sun, Wen, Wang, Zixuan, Gao, Xinmeng, Cao, Zhenyuan, Ouyang, Yusheng, Yao, Lang, Wang, Yifan, Xiao, Yunqi, Li, Ruidong, An, Xingwang, Dai, Xinyi, Zhao, Lulu, Qian, Lu, Zhang, Panke, Huang, Shuo

Issue&Volume: 2026-07-29

Abstract: Accurate decoding of peptide sequences is crucial in proteomics. However, achieving this goal is a technical challenge, owing to the compositional and structural complexity of peptides1. Studies inspired by the success of nanopore nucleic acid sequencing have shown that nanopore-based techniques can also be applied to peptide sequencing2,3. The key is to generate narrowly distributed, consistent and sequence-dependent events during sequential nanopore readout. Here we introduce a nanopore-based strategy termed transient pore analyte looping (tPAL). We develop an engineered Mycobacterium smegmatis porin A (MspA) nanopore that is dual modified with a nickel-ion-bound nitrilotriacetic acid (NTA-Ni) adapter and the target peptide. This distinctive sensing configuration enables precise recognition of the Nterminus of the immobilized peptide by multiple re-readings. With the aid of cholesterolized aminopeptidase, the immobilized peptide can be shortened sequentially in single-amino-acid increments, yielding sequence-dependent, stepwise and narrowly distributed signal alterations that provide clues to allow peptide sequence decoding. Our strategy achieves single-amino-acid resolution and effectively identifies single-amino-acid mutations, post-translational modifications and unnatural-amino-acid insertions—indicative of its versatility in nanopore proteomics and chiral peptide analyses.

DOI: 10.1038/s41586-026-10881-1

Source: https://www.nature.com/articles/s41586-026-10881-1