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首页> 外文期刊>European Journal of Cell Biology: Journal of Deutsche Gesellschaft fur Elektronenmikroskopie: Journal of Deutsche Gesellschaft fur Zellbiologie >Tyrosine phosphorylation of 40 kDa proteins in osteoblastic cells aftermechanical stimulation of beta(1)-integrins
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Tyrosine phosphorylation of 40 kDa proteins in osteoblastic cells aftermechanical stimulation of beta(1)-integrins

机译:机械刺激β(1)-整合素后成骨细胞中40 kDa蛋白的酪氨酸磷酸化。

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摘要

Using a method for the mechanical stimulation of cells which was adapted from one developed by Wang and Ingber employing magnetic microbeads [Wang, N. D., D. E. Ingber: Control of cytoskeletal mechanics by extracellular matrix, cell shape, and mechanical tension, Biophys, J. 66, 2181-2189 (1994)], mechanical stress could be applied to specific receptors on the cell surface. To achieve this, ferromagnetic microbeads coated with different ligands were magnetized after adhesion to the cells, The beads were then 'twisted' using a second magnetic field oriented perpendicular to the magnetizing one, Contrary to most current methods, it was possible to confer the strain without deforming the cell as a whole, thus being able to observe the individual reactions of transmembrane receptors to mechanical stress. An increase in tyrosine phosphorylation of proteins migrating at approximately 40 kDa could be observed as a reaction to stress on the beta(1)-subunits of the integrin family, while stress to other transmembrane molecules like the transferrin or low density lipoprotein receptors with no connection to the cytoskeleton did not give this reaction, Fibroblastic cells showed, contrary to osteoblastic cells, no reaction to stress applied on transmembrane proteins.
机译:使用一种由Wang和Ingber开发的方法对细胞进行机械刺激的方法,该方法采用了磁微珠[Wang,ND,DE Ingber:通过细胞外基质,细胞形状和机械张力控制细胞骨架力学,Biophys,J. 66 ,2181-2189(1994)],可以将机械应力施加到细胞表面的特定受体上。为此,将涂有不同配体的铁磁微珠粘附到细胞后进行磁化。然后使用垂直于磁化磁场的第二磁场“扭曲”磁珠,与大多数当前方法相反,可以赋予应变不会使整个细胞变形,因此能够观察跨膜受体对机械应力的个别反应。观察到对整合素家族β(1)-亚基的应激反应,蛋白质迁移约40 kDa时酪氨酸磷酸化增加,而对其他跨膜分子(如转铁蛋白或低密度脂蛋白受体)的应激则没有关系与成骨细胞相反,成纤维细胞表明,对细胞骨架没有反应,对成膜细胞的应力没有反应。

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