Mutoh T, Naoi M, Sobue I, Kiuchi K, Nagatsu T
Clin Chim Acta. 1985 Nov 15;152(3):307-14. doi: 10.1016/0009-8981(85)90106-8.
To study the enzymatic properties of beta-galactosidase from the patients with a beta-galactosidase deficiency such as GM1 gangliosidosis, determination of enzymatic activity with naturally occurring substrates, asialofetuin in addition to another natural substrate, GM1 ganglioside, is essentially required. With a previously reported, simple and sensitive fluorometric assay for GM1 ganglioside beta-galactosidase using high performance liquid chromatography (HPLC), optimal reaction conditions were determined for the assay of acid beta-galactosidase activity toward asialofetuin in skin fibroblast homogenates. Under these conditions, reduced enzymatic activities could be detected in cultured skin fibroblasts from patients with type 1 and 3 GM1 gangliosidoses and mucopolysaccharidosis IV-B (Morquio B syndrome). This method was applicable to study of the enzymatic properties of the mutant beta-galactosidase and provided an alternative to assays employing radioactive or artificial substrates.
为研究患有β-半乳糖苷酶缺乏症（如GM1神经节苷脂贮积症）患者的β-半乳糖苷酶的酶学特性，除了另一种天然底物GM1神经节苷脂外，使用天然存在的底物脱唾液酸胎球蛋白测定酶活性是必不可少的。利用先前报道的、使用高效液相色谱（HPLC）对GM1神经节苷脂β-半乳糖苷酶进行的简单且灵敏的荧光测定法，确定了用于测定皮肤成纤维细胞匀浆中酸性β-半乳糖苷酶对脱唾液酸胎球蛋白活性的最佳反应条件。在这些条件下，可在1型和3型GM1神经节苷脂贮积症患者以及黏多糖贮积症IV-B（莫尔基奥B综合征）患者的培养皮肤成纤维细胞中检测到酶活性降低。该方法适用于研究突变型β-半乳糖苷酶的酶学特性，并为采用放射性或人工底物的测定提供了一种替代方法。
