Belyy V, Yildiz A
Biophysics Graduate Group, University of California at Berkeley, Berkeley, CA, United States.
University of California at Berkeley, Berkeley, CA, United States.
Methods Enzymol. 2017;582:31-54. doi: 10.1016/bs.mie.2016.09.037. Epub 2016 Dec 3.
Cytoskeletal motors utilize the energy stored in ATP to generate linear motion along rigid filaments. Because their enzymatic cycles are tightly coupled to the production of force and forward movement, the optical-trapping technique is uniquely suited for studying their mechanochemical cycle. Here, we discuss the practical aspects of optical trapping in connection with single-motor assays and describe three distinct experimental modes (fixed-trap, force feedback, and square wave) that are typically used to investigate the enzymatic and biophysical properties of cytoskeletal motors. The principal outstanding questions in the field involve motor regulation by cargo adaptor proteins and cargo transport by teams of motors, ensuring that the optical trap's ability to apply precise forces and measure nanometer-scale displacements will remain crucial to the study of intracellular motility in the foreseeable future.
细胞骨架马达利用储存在三磷酸腺苷（ATP）中的能量，沿着刚性细丝产生线性运动。由于它们的酶促循环与力的产生和向前运动紧密耦合，光镊技术特别适合于研究它们的机械化学循环。在这里，我们讨论与单马达分析相关的光镊实际应用，并描述三种不同的实验模式（固定陷阱、力反馈和方波），这些模式通常用于研究细胞骨架马达的酶学和生物物理特性。该领域主要的突出问题涉及货物衔接蛋白对马达的调节以及马达团队的货物运输，这确保了在可预见的未来，光镊施加精确力和测量纳米级位移的能力对于细胞内运动性的研究仍然至关重要。
