小柯机器人

南京大学Shuo Huang小组在研究中取得进展。他们的最新研究利用纳米孔无偏倚区分了20种蛋白质氨基酸及其修饰。2023年9月25日出版的《自然-方法学》发表了这项成果。

研究人员建立一种工程异质八聚体包皮分枝杆菌porin A（MspA）纳米孔，其中包含唯一的Ni²⁺修饰。它可以完全区分所有20种蛋白氨基酸和4种代表性修饰氨基酸，Nω、N'ω-二甲基精氨酸（Me-R）、O-乙酰苏氨酸（Ac-T）、N4-（β-N-乙酰基-葡萄糖胺基）-天冬酰胺（GlcNAc-N）和O-磷丝质碱（P-S）。在机器学习的帮助下，其准确率达到了98.6%。

研究人员还使用该工具分析了氨基酸补充片剂和肽酶消化的氨基酸。这种同时区分所有20种蛋白氨基酸及其翻译后修饰（PTM）的能力，表明该纳米孔测序方法在蛋白质测序中的巨大应用潜能。

据悉，天然蛋白质由20种蛋白质氨基酸及其翻译后修饰组成。然而，由于缺乏合适的纳米孔传感器可以同时区分所有20种氨基酸及其PTM，因此尚未实现利用纳米孔对蛋白质进行直接测序。

附：英文原文

Title: Unambiguous discrimination of all 20 proteinogenic amino acids and their modifications by nanopore

Author: Wang, Kefan, Zhang, Shanyu, Zhou, Xiao, Yang, Xian, Li, Xinyue, Wang, Yuqin, Fan, Pingping, Xiao, Yunqi, Sun, Wen, Zhang, Panke, Li, Wenfei, Huang, Shuo

Issue&Volume: 2023-09-25

Abstract: Natural proteins are composed of 20 proteinogenic amino acids and their post-translational modifications (PTMs). However, due to the lack of a suitable nanopore sensor that can simultaneously discriminate between all 20 amino acids and their PTMs, direct sequencing of protein with nanopores has not yet been realized. Here, we present an engineered hetero-octameric Mycobacterium smegmatis porin A (MspA) nanopore containing a sole Ni2+ modification. It enables full discrimination of all 20 proteinogenic amino acids and 4 representative modified amino acids, Nω,N’ω-dimethyl-arginine (Me-R), O-acetyl-threonine (Ac-T), N4-(β-N-acetyl-D-glucosaminyl)-asparagine (GlcNAc-N) and O-phosphoserine (P-S). Assisted by machine learning, an accuracy of 98.6% was achieved. Amino acid supplement tablets and peptidase-digested amino acids from peptides were also analyzed using this strategy. This capacity for simultaneous discrimination of all 20 proteinogenic amino acids and their PTMs suggests the potential to achieve protein sequencing using this nanopore-based strategy.

DOI: 10.1038/s41592-023-02021-8

Source: https://www.nature.com/articles/s41592-023-02021-8

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex