Construction and operation of high-resolution magnetic tape head tweezers for measuring single-protein dynamics under force.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology
Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-02-05 DOI:10.1016/bs.mie.2023.12.010
Rafael Tapia-Rojo
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引用次数: 0

Abstract

Mechanical forces are critical to protein function across many biological contexts-from bacterial adhesion to muscle mechanics and mechanotransduction processes. Hence, understanding how mechanical forces govern protein activity has developed into a central scientific question. In this context, single-molecule magnetic tweezers has recently emerged as a valuable experimental tool, offering the capability to measure single proteins over physiologically relevant forces and timescales. In this chapter, we present a detailed protocol for the assembly and operation of our magnetic tape head tweezers instrument, specifically tailored to investigate protein dynamics. Our instrument boasts a simplified microscope design and incorporates a magnetic tape head as the force-generating apparatus, facilitating precise force control and enhancing its temporal stability, enabling the study of single protein mechanics over extended timescales spanning several hours or even days. Moreover, its straightforward and cost-effective design ensures its accessibility to the wider scientific community. We anticipate that this technique will attract widespread interest within the growing field of mechanobiology and expect that this chapter will provide facilitated accessibility to this technology.

用于测量单个蛋白质受力动态的高分辨率磁带头镊子的构造和操作。
从细菌粘附到肌肉力学和机械传导过程,机械力对许多生物环境中的蛋白质功能至关重要。因此,了解机械力如何支配蛋白质的活动已成为一个核心科学问题。在这种情况下,单分子磁镊最近成为一种有价值的实验工具,能够测量单个蛋白质的生理相关力和时间尺度。在本章中,我们将详细介绍磁带头镊子仪器的组装和操作步骤,该仪器专门用于研究蛋白质动力学。我们的仪器采用了简化的显微镜设计,并将磁带头作为发力装置,从而实现了精确的力控制,并增强了其时间稳定性,从而能够在数小时甚至数天的较长时间尺度内研究单个蛋白质的力学。此外,它的设计简单明了、成本效益高,确保了更广泛的科学界可以使用它。我们预计,这项技术将在不断发展的机械生物学领域引起广泛兴趣,并希望本章能为这项技术的应用提供便利。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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