Dahl D, Bignami A
Biochim Biophys Acta. 1979 Jun 19;578(2):305-16. doi: 10.1016/0005-2795(79)90161-2.
The glial fibrillary acidic protein and an immunologically active cyanogen bromide peptide were purified by immunoaffinity chromatography from 8 M urea extracts of brain filament preparations isolated from bovine white matter according to Norton's procedure. The protein accounted for approximately 30% of the total protein in this preparation and for the largest fraction in the 50 000 molecular weight range. The fraction not absorbed to the immuno-Sepharose column reacted with neurofilament antisera by double immunodiffusion. On sodium dodecyl sulfate gel electrophoresis the main bands in the non-adsorbed fraction were at 74 000 daltons and above 100 000. Several bands were seen in the 50 000 molecular weight range. It is concluded that glio- and neurofilaments co-purify together in Norton's procedure and that neurofilaments are probably heterogeneous in polypeptide composition.
根据诺顿的方法,从牛白质分离出的脑丝状体制剂的8M尿素提取物中,通过免疫亲和层析法纯化了胶质纤维酸性蛋白和一种具有免疫活性的溴化氰肽。该蛋白约占该制剂总蛋白的30%,是50000分子量范围内最大的组分。未被免疫琼脂糖柱吸附的部分通过双向免疫扩散与神经丝抗血清发生反应。在十二烷基硫酸钠凝胶电泳上,未吸附部分的主要条带位于74000道尔顿及100000道尔顿以上。在50000分子量范围内可见几条条带。得出的结论是,在诺顿的方法中,胶质丝和神经丝共同纯化,并且神经丝在多肽组成上可能是异质的。