建立生物原子力显微镜以研究神经退行性疾病中的蛋白质错误折叠。

Setting Up a Bio-AFM to Study Protein Misfolding in Neurodegenerative Diseases.

机构信息

Bioinformatics and Human Electrophysiology Laboratory, Department of Informatics, Ionian University, Corfu, Greece.

出版信息

Adv Exp Med Biol. 2023;1423:1-10. doi: 10.1007/978-3-031-31978-5_1.

DOI:10.1007/978-3-031-31978-5_1

Abstract

The clinical pathology of neurodegenerative diseases suggests that earlier onset and progression are related to the accumulation of protein aggregates due to misfolding. A prominent way to extract useful information regarding single-molecule studies of protein misfolding at the nanoscale is by capturing the unbinding molecular forces through forced mechanical tension generated and monitored by an atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS). This AFM-driven process results in an amount of data in the form of force versus molecular extension plots (force-distance curves), the statistical analysis of which can provide insights into the underlying energy landscape and assess a number of characteristic elastic and kinetic molecular parameters of the investigated sample. This chapter outlines the setup of a bio-AFM-based SMFS technique for single-molecule probing. The infrastructure used as a reference for this presentation is the Bruker ForceRobot300.

摘要

神经退行性疾病的临床病理学表明，早期发病和进展与蛋白质聚集物的积累有关，这些聚集物是由于错误折叠而产生的。在纳米尺度上研究蛋白质错误折叠的单分子研究中，一种突出的方法是通过原子力显微镜（AFM）单分子力谱（AFM-SMFS）来捕获未结合的分子力，这种方法可以通过产生和监测强制机械张力来实现。这个 AFM 驱动的过程会产生大量的数据，形式为力与分子延伸图（力-距离曲线），对其进行统计分析可以深入了解潜在的能量景观，并评估研究样本的许多特征弹性和动力学分子参数。本章概述了基于生物 AFM 的单分子探测 SMFS 技术的设置。本演示文稿所使用的基础设施是 Bruker ForceRobot300。