Ju Lining, Chen Yunfeng, Xue Lingzhou, Du Xiaoping, Zhu Cheng
Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, United States.
Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, United States.
Elife. 2016 Jul 19;5:e15447. doi: 10.7554/eLife.15447.
How cells sense their mechanical environment and transduce forces into biochemical signals is a crucial yet unresolved question in mechanobiology. Platelets use receptor glycoprotein Ib (GPIb), specifically its α subunit (GPIbα), to signal as they tether and translocate on von Willebrand factor (VWF) of injured arterial surfaces against blood flow. Force elicits catch bonds to slow VWF-GPIbα dissociation and unfolds the GPIbα leucine-rich repeat domain (LRRD) and juxtamembrane mechanosensitive domain (MSD). How these mechanical processes trigger biochemical signals remains unknown. Here we analyze these extracellular events and the resulting intracellular Ca(2+) on a single platelet in real time, revealing that LRRD unfolding intensifies Ca(2+) signal whereas MSD unfolding affects the type of Ca(2+) signal. Therefore, LRRD and MSD are analog and digital force transducers, respectively. The >30 nm macroglycopeptide separating the two domains transmits force on the VWF-GPIbα bond (whose lifetime is prolonged by LRRD unfolding) to the MSD to enhance its unfolding, resulting in unfolding cooperativity at an optimal force. These elements may provide design principles for a generic mechanosensory protein machine.
细胞如何感知其力学环境并将力转化为生化信号,是力学生物学中一个关键但尚未解决的问题。血小板利用受体糖蛋白Ib(GPIb),特别是其α亚基(GPIbα),在逆着血流在受损动脉表面的血管性血友病因子(VWF)上拴系和移位时发出信号。力引发捕捉键以减缓VWF-GPIbα解离,并使GPIbα富含亮氨酸的重复结构域(LRRD)和近膜机械敏感结构域(MSD)展开。这些机械过程如何触发生化信号仍然未知。在这里,我们实时分析了单个血小板上的这些细胞外事件以及由此产生的细胞内Ca(2+),揭示了LRRD展开增强Ca(2+)信号,而MSD展开影响Ca(2+)信号的类型。因此,LRRD和MSD分别是模拟和数字力传感器。分隔这两个结构域的>30 nm大糖肽将VWF-GPIbα键(其寿命因LRRD展开而延长)上的力传递给MSD以增强其展开,从而在最佳力下产生展开协同性。这些元件可能为通用的机械传感蛋白机器提供设计原则。