Kreplak Laurent, Herrmann Harald, Aebi Ueli
M. E Müller Institute for Structural Biology, Biozentrum, University of Basel, 4056 Basel, Switzerland.
Biophys J. 2008 Apr 1;94(7):2790-9. doi: 10.1529/biophysj.107.119826. Epub 2008 Jan 4.
Within muscle fibers, desmin intermediate filaments (IFs) are major constituents of the extrasarcomeric cytoskeleton. However, their contribution to the mechanical properties of myocytes has remained elusive. We present an experimental approach to measure the extensibility and the tensile strength of in vitro reconstituted desmin IFs adsorbed to a solid support. The tip of an atomic force microscope (AFM) was used to push on single filaments perpendicular to the filament axis. The torque of the AFM cantilever was monitored during the pushing events to yield an estimate of the lateral force necessary to bend and stretch the filaments. Desmin IFs were stretched up to 3.4-fold with a maximum force of approximately 3.5 nN. Fully stretched filaments exhibited a much smaller diameter than did native IFs, i.e., approximately 3.5 nm compared to 12.6 nm, both by AFM and electron microscopy. Moreover, we combined the morphological and lateral force data to compute an average stress-strain curve for a single desmin filament. The main features were a pronounced strain-hardening regime above 50% extension and a tensile strength of at least 240 MPa. Because of these nonlinear tensile properties, desmin IFs may dissipate mechanical energy and serve as a physical link between successive sarcomeres during large deformation.
在肌纤维内,结蛋白中间丝(IFs)是肌节外细胞骨架的主要成分。然而,它们对心肌细胞力学特性的贡献仍不明确。我们提出了一种实验方法,用于测量吸附在固体支持物上的体外重构结蛋白IFs的可伸展性和拉伸强度。使用原子力显微镜(AFM)的尖端垂直于细丝轴推压单根细丝。在推压过程中监测AFM悬臂的扭矩,以估算弯曲和拉伸细丝所需的侧向力。结蛋白IFs可拉伸至3.4倍,最大力约为3.5 nN。通过AFM和电子显微镜观察,完全拉伸的细丝直径比天然IFs小得多,即分别约为3.5 nm和12.6 nm。此外,我们结合形态学和侧向力数据,计算了单个结蛋白细丝的平均应力-应变曲线。主要特征是在延伸率超过50%时出现明显的应变硬化阶段,拉伸强度至少为240 MPa。由于这些非线性拉伸特性,结蛋白IFs可能耗散机械能,并在大变形过程中作为连续肌节之间的物理连接。