James G T, Thach A B, Klassen L, Austin J H
Life Sci. 1985 Dec 23;37(25):2365-71. doi: 10.1016/0024-3205(85)90103-1.
In this report we describe a method to purify both normal and abnormal (inactive) arylsulfatase A. The abnormal enzyme protein was isolated both from cases of late infantile and early juvenile forms of metachromatic leukodystrophy. Conventional protein isolation methods reported earlier were followed by size exclusion high-performance liquid chromatography in the final purification stages. Both the mutant enzyme and the normal enzyme had the same HPLC elution behavior. They thus appeared to self-associate in a similar pH-dependent fashion. Both could be followed by their reaction to a rabbit antibody to normal human arylsulfatase A. The amount of homogenous protein obtained from about 500 grams of liver was 300-400 micrograms.
在本报告中，我们描述了一种纯化正常和异常（无活性）芳基硫酸酯酶A的方法。异常酶蛋白是从晚期婴儿型和早期青少年型异染性脑白质营养不良病例中分离出来的。在最后的纯化阶段，先采用早期报道的传统蛋白质分离方法，然后进行尺寸排阻高效液相色谱法。突变酶和正常酶具有相同的高效液相色谱洗脱行为。因此，它们似乎以类似的pH依赖性方式进行自我缔合。两者都可以通过它们与兔抗正常人芳基硫酸酯酶A抗体的反应来追踪。从约500克肝脏中获得的纯蛋白量为300 - 400微克。
