Fressinaud C, Sarliève L L, Labourdette G
Centre de Neurochimie, C.N.R.S., Strasbourg.
C R Acad Sci III. 1990;310(7):303-9.
We describe a method of partial purification of rat brain 3'-phosphoadenosine-5'-phosphosulfate galactocerebroside sulfotransferase (EC 2.8.2.11, CST). The first steps consist of a Triton X-100 extraction of the enzyme from delipidated microsomes and the fractionation of this extract by precipitation with ammonium sulfate, followed by successive chromatography on hydrophobic gel (phenyl sepharose), heparin, gel filtration (Trisacryl GF2000), hydroxy-apatite and cation exchange resin (SP trisacryl). The denaturation of the enzyme and its instability account for the low enrichment achieved in terms of specific activity, but an evident simplification of the electrophoretic pattern was obtained.
我们描述了一种大鼠脑3'-磷酸腺苷-5'-磷酸硫酸半乳糖脑苷脂磺基转移酶(EC 2.8.2.11,CST)的部分纯化方法。第一步包括用 Triton X-100 从脱脂微粒体中提取该酶,然后通过硫酸铵沉淀对该提取物进行分级分离,接着依次在疏水凝胶(苯基琼脂糖)、肝素、凝胶过滤(Trisacryl GF2000)、羟基磷灰石和阳离子交换树脂(SP trisacryl)上进行层析。该酶的变性及其不稳定性导致在比活性方面获得的富集程度较低,但电泳图谱明显简化。