荷兰格罗宁根大学教授Maglia, Giovanni团队自下而上制造蛋白酶体-纳米孔分解和处理单个蛋白质。相关研究成果于2021年11月18日发表在《自然—化学》。

在大多数单分子方法中，能够处理生物分子的分子机器的精确组装和工程设计起着至关重要的作用。

该文中，研究人员使用从生命的所有三个领域的组成部分，制造了一个集成的多蛋白复合物，该复合物控制单个蛋白质在纳米孔上的展开和线程。该900kDa多组分设备分两步制造。首先，通过将细菌保护性抗原的跨膜区连接到哺乳动物蛋白酶体激活剂，设计了一种稳定、低噪声的β-桶纳米孔传感器。然后将一个古老的20S蛋白酶体构建到人工纳米孔中，以控制蛋白质在纳米孔中的展开和线性运输。

该多组分分子机器为单分子蛋白质分析中的两种方法打开了大门，在该方法中，选定的底物蛋白质被展开，送入蛋白酶体室，然后作为片段肽或完整多肽进行处理。

附：英文原文

Title: Bottom-up fabrication of a proteasome–nanopore that unravels and processes single proteins

Author: Zhang, Shengli, Huang, Gang, Versloot, Roderick Corstiaan Abraham, Bruininks, Bart Marlon Herwig, de Souza, Paulo Cesar Telles, Marrink, Siewert-Jan, Maglia, Giovanni

Issue&Volume: 2021-11-18

Abstract: The precise assembly and engineering of molecular machines capable of handling biomolecules play crucial roles in most single-molecule methods. In this work we use components from all three domains of life to fabricate an integrated multiprotein complex that controls the unfolding and threading of individual proteins across a nanopore. This 900kDa multicomponent device was made in two steps. First, we designed a stable and low-noise β-barrel nanopore sensor by linking the transmembrane region of bacterial protective antigen to a mammalian proteasome activator. An archaeal 20S proteasome was then built into the artificial nanopore to control the unfolding and linearized transport of proteins across the nanopore. This multicomponent molecular machine opens the door to two approaches in single-molecule protein analysis, in which selected substrate proteins are unfolded, fed to into the proteasomal chamber and then addressed either as fragmented peptides or intact polypeptides.

DOI: 10.1038/s41557-021-00824-w

Source: https://www.nature.com/articles/s41557-021-00824-w