Stigbrand T, Eriksson A, Thornell L E
Biochim Biophys Acta. 1979 Mar 27;577(1):52-60. doi: 10.1016/0005-2795(79)90007-2.
The intermediate filament protein skeletin from cow heart Purkinje fibres was purified to homogeneity by a selective extraction procedure and gel chromatography in the presence of sodium dodecyl sulphate. Monospecific antibodies were obtained by immunisation of rabbits with the sodium dodecyl sulphate-skeletin complex, and rocket electrophoresis made it possible to quantify the concentration of protein. The skeletin monomer has a molecular weight of 55 000. Amino acid analysis revealed that skeletin has a high content of glutamic acid, aspartic acid, alanine and leucine, together constituting more than 50% of the molecule. The isoelectric point is determined as 6.35. Skeletin is insoluble at pH 4--6 in the absence of detergent and shows increasing solubility at higher and lower pH. The biochemical characteristics are discussed in relation to the cytoskeletal function of the filaments. Comparison with intermediate-sized filament protein of other tissues show certain important similarities suggesting that the filaments may share a common evolutionary ancestry.
通过一种选择性提取方法以及在十二烷基硫酸钠存在下的凝胶色谱法，从牛心脏浦肯野纤维中纯化出了中间丝蛋白骨骼蛋白，使其达到了均一性。用十二烷基硫酸钠 - 骨骼蛋白复合物免疫兔子获得了单特异性抗体，火箭电泳使得对蛋白质浓度进行定量成为可能。骨骼蛋白单体的分子量为55000。氨基酸分析表明，骨骼蛋白含有高含量的谷氨酸、天冬氨酸、丙氨酸和亮氨酸，它们总共构成了分子的50%以上。其等电点测定为6.35。在没有去污剂的情况下，骨骼蛋白在pH 4 - 6时不溶，而在更高和更低的pH值下溶解度增加。结合细丝的细胞骨架功能对其生化特性进行了讨论。与其他组织的中间丝蛋白进行比较显示出某些重要的相似性，这表明这些细丝可能有着共同的进化起源。
