Scheibe R, Wenzel K W, Hofmann E
Biomed Biochim Acta. 1985;44(9):1279-86.
Acid alpha-glucosidase was purified from the lysosomal/mitochondrial fraction of rat liver by acid precipitation, ammonium sulphate precipitation and affinity chromatography on Sephadex G 100, resulting in 17000-fold enrichment from that of the liver homogenate. The molecular weight of the enzyme was estimated to be 125000 from analytical gel filtration experiments. On SDS-polyacrylamide gel electrophoresis the purified enzyme showed only a single band having an apparent molecular weight of about 64000. Based on these results, it is concluded that lysosomal liver alpha-glucosidase consists of two subunits. The discontinuous system of SDS-polyacrylamide gel electrophoresis, however, revealed two closely migrating protein bands suggesting heterogeneity of the enzyme subunits.
酸性α-葡萄糖苷酶通过酸沉淀、硫酸铵沉淀以及在葡聚糖凝胶G 100上进行亲和层析，从大鼠肝脏的溶酶体/线粒体组分中纯化得到，相对于肝脏匀浆实现了17000倍的富集。通过分析性凝胶过滤实验估计该酶的分子量为125000。在SDS-聚丙烯酰胺凝胶电泳中，纯化后的酶仅显示出一条表观分子量约为64000的单一谱带。基于这些结果，可以得出结论，溶酶体肝脏α-葡萄糖苷酶由两个亚基组成。然而，SDS-聚丙烯酰胺凝胶电泳的不连续系统显示出两条迁移紧密的蛋白谱带，表明该酶亚基具有异质性。
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