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.
摘要:
从细菌粘附到肌肉力学和机械转导过程,机械力对于许多生物学环境中的蛋白质功能至关重要。因此,理解机械力如何控制蛋白质活性已经发展成为一个核心科学问题。在这种情况下,单分子磁镊子最近已经成为一种有价值的实验工具,提供在生理相关的力量和时间尺度上测量单个蛋白质的能力。在这一章中,我们为我们的磁带头镊子仪器的组装和操作提供了详细的协议,专门为研究蛋白质动力学而定制。我们的仪器拥有简化的显微镜设计,并采用磁带头作为力产生装置,促进精确的力控制并增强其时间稳定性,能够在几个小时甚至几天的延长时间尺度上研究单一蛋白质力学。此外,其直截了当且具有成本效益的设计确保了其对更广泛的科学界的可及性。我们预计该技术将在不断发展的机械生物学领域引起广泛的兴趣,并希望本章将为该技术提供便利的可访问性。