Inagaki M, Nishi Y, Nishizawa K, Matsuyama M, Sato C
Nature. 1987;328(6131):649-52. doi: 10.1038/328649a0.
Intermediate filaments are a major component of the cytoskeleton of eukaryotic cells. Although there appear to be at least five distinct classes of these filaments, cells of mesenchymal origin and most cells in culture contain the intermediate filament composed of the subunit protein vimentin. Vimentin exists in a nonphosphorylated as well as in a phosphorylated form, and there is increased phosphorylation of this protein when the filament undergoes marked redistribution in various cells. The role of phosphorylation on assembly-disassembly and organization of the vimentin filament has remained obscure. We report here a stable and purified system allowing biochemical examination of vimentin filament assembly and disassembly. Using this in vitro system, we carried out stoichiometrical phosphorylations, using purified protein kinases. We obtained evidence for site-specific, phosphorylation-dependent disassembly of the vimentin filament.
中间丝是真核细胞细胞骨架的主要组成部分。尽管这些丝似乎至少有五个不同的类别，但间充质来源的细胞和大多数培养细胞含有由亚基蛋白波形蛋白组成的中间丝。波形蛋白以非磷酸化形式和磷酸化形式存在，当丝在各种细胞中经历明显的重新分布时，这种蛋白的磷酸化会增加。磷酸化在波形蛋白丝的组装 - 拆卸和组织中的作用仍然不清楚。我们在此报告一个稳定且纯化的系统，可用于对波形蛋白丝的组装和拆卸进行生化检查。使用这个体外系统，我们使用纯化的蛋白激酶进行了化学计量的磷酸化。我们获得了波形蛋白丝位点特异性、磷酸化依赖性拆卸的证据。
