Evans R M
Department of Pathology, University of Colorado Health Sciences Center, Denver 80262.
Eur J Cell Biol. 1988 Apr;46(1):152-60.
Analysis of specific fragments of vimentin and desmin from 32P-labeled BHK-21 cells indicated that these intermediate-filament subunit proteins are phosphorylated in specific regions or domains. High performance liquid chromatography and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis of lysine-specific protease-generated fragments demonstrated that both molecules were phosphorylated in their amino terminal or "head" domains. While this was the predominant site of phosphorylation for vimentin, additional phosphorylated fragments from desmin were observed. Chemical cleavage of [32P]desmin and subsequent examination of the phosphorylated peptides indicated that the major site of desmin phosphorylation was located within the "tail" domain. Analysis of vimentin and desmin from non-mitotic and mitotically selected cells indicated that the increased phosphorylation of intermediate-filament proteins observed during cell division occurs within the amino terminal domains of both molecules. These studies indicate that the increased phosphorylation of filament proteins during mitosis may involve the function of the amino terminal domain. In addition, filament proteins may be phosphorylated in a subunit-protein-specific manner which may reflect subunit-specific functions.
对来自经³²P标记的BHK - 21细胞的波形蛋白和结蛋白的特定片段进行分析表明,这些中间丝亚基蛋白在特定区域或结构域发生了磷酸化。对赖氨酸特异性蛋白酶产生的片段进行高效液相色谱和十二烷基硫酸钠(SDS)聚丙烯酰胺凝胶电泳显示,这两种分子在其氨基末端或“头部”结构域都发生了磷酸化。虽然这是波形蛋白磷酸化的主要位点,但观察到结蛋白还有其他磷酸化片段。对[³²P]结蛋白进行化学裂解并随后检查磷酸化肽段表明,结蛋白磷酸化的主要位点位于“尾部”结构域。对非有丝分裂和经有丝分裂选择的细胞中的波形蛋白和结蛋白进行分析表明,在细胞分裂过程中观察到的中间丝蛋白磷酸化增加发生在这两种分子的氨基末端结构域内。这些研究表明,有丝分裂期间丝蛋白磷酸化增加可能涉及氨基末端结构域的功能。此外,丝蛋白可能以亚基蛋白特异性的方式发生磷酸化,这可能反映了亚基特异性的功能。