Wang C, Asai D J, Lazarides E
Proc Natl Acad Sci U S A. 1980 Mar;77(3):1541-5. doi: 10.1073/pnas.77.3.1541.
Purified preparations of 10-nm neurofilaments from rat spinal cord and bovine or porcine brain contain a predominant 68,000-dalton polypeptide. This polypeptide is also a major component of the neurofilaments that copurify with brain tubulin isolated by cycles of polymerization and depolymerization. A protein that has the same isoelectric point and molecular weight as the neurofilament-associated polypeptide has also been identified as a cytoskeletal protein in a variety of avian and mammalian cell types, including baby hamster kidney (BHK-21) mouse 3T3, Novikoff rat hepatoma, chicken fibroblast, and chicken muscle cells. This protein is also a component of isolated chicken skeletal myofibrils. One-dimensional peptide maps of the 68,000-dalton proteins purified by two-dimensional isoelectric focusing/NaDodSO(4)/polyacrylamide gel electrophoresis from myofibrils, cycled tubulin, purified neurofilaments, and various cultured cell types were identical. In immunofluorescence this protein was associated with cytoplasmic intermediate filaments and myofibril Z discs. These results indicate that the neurofilament-associated polypeptide is a conserved protein that is present in many different cell types in addition to neuronal cells. Because some of these cells contain the major components of two other intermediate filament classes, desmin and vimentin, a given cell type may contain the subunits of at least three distinct intermediate filament types.
从大鼠脊髓以及牛或猪脑中提纯的10纳米神经丝制剂含有一种占主导地位的68000道尔顿多肽。这种多肽也是通过聚合和解聚循环分离出的与脑微管蛋白共纯化的神经丝的主要成分。一种与神经丝相关多肽具有相同等电点和分子量的蛋白质，在多种鸟类和哺乳动物细胞类型中也被鉴定为细胞骨架蛋白，包括幼仓鼠肾（BHK - 21）细胞、小鼠3T3细胞、诺维科夫大鼠肝癌细胞、鸡成纤维细胞和鸡肌肉细胞。这种蛋白质也是分离出的鸡骨骼肌肌原纤维的组成成分。通过二维等电聚焦/十二烷基硫酸钠/聚丙烯酰胺凝胶电泳从肌原纤维、循环微管蛋白、纯化的神经丝以及各种培养细胞类型中纯化出的68000道尔顿蛋白质的一维肽图是相同的。在免疫荧光中，这种蛋白质与细胞质中间丝和肌原纤维Z盘相关。这些结果表明，神经丝相关多肽是一种保守蛋白，除神经元细胞外，还存在于许多不同的细胞类型中。由于其中一些细胞含有另外两种中间丝类型（结蛋白和波形蛋白）的主要成分，所以特定的细胞类型可能包含至少三种不同中间丝类型的亚基。
