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科学家利用纳米孔在单氨基酸分辨率下对单一蛋白质进行读取
作者:小柯机器人 发布时间:2021/11/7 13:15:15

荷兰代尔夫特理工大学Cees Dekker团队利用纳米孔在单氨基酸分辨率下对单一蛋白质进行读取。2021年11月4日,《科学》杂志在线发表了这项成果。

研究人员展示了一种基于纳米孔的单分子肽阅读器,它对单个肽内的单氨基酸替换敏感。一个DNA-肽共轭物被DNA螺旋酶Hel308拉过生物纳米孔MspA。通过纳米孔读取离子电流信号,可以辨别单个读数中的单氨基酸替换。分子动力学模拟显示这些信号是由大小排异和孔隙结合造成的。

研究人员还展示了"倒退"肽读数的能力,获得了同一分子的许多独立读数,并在单氨基酸变体鉴定中产生的错误率<10-6。这些概念验证实验构成了开发单分子蛋白质指纹和分析技术的基础。

据介绍,能够识别单个蛋白质的蛋白组学工具对细胞生物学研究和应用非常重要。

附:英文原文

Title: Multiple rereads of single proteins at single–amino acid resolution using nanopores

Author: Henry Brinkerhoff, Albert S. W. Kang, Jingqian Liu, Aleksei Aksimentiev, Cees Dekker

Issue&Volume: 2021-11-04

Abstract: A proteomics tool capable of identifying single proteins would be important for cell biology research and applications. Here, we demonstrate a nanopore-based single-molecule peptide reader sensitive to single–amino acid substitutions within individual peptides. A DNA-peptide conjugate was pulled through the biological nanopore MspA by the DNA helicase Hel308. Reading the ion current signal through the nanopore enabled discrimination of single–amino acid substitutions in single reads. Molecular dynamics simulations showed these signals to result from size exclusion and pore binding. We also demonstrate the capability to “rewind” peptide reads, obtaining numerous independent reads of the same molecule, yielding an error rate <106 in single amino acid variant identification. These proof-of-concept experiments constitute a promising basis for the development of a single-molecule protein fingerprinting and analysis technology.

DOI: abl4381

Source: https://www.science.org/doi/10.1126/science.abl4381

 

期刊信息
Science:《科学》,创刊于1880年。隶属于美国科学促进会,最新IF:41.037