Choy F Y, Woo M, Potier M
Biochim Biophys Acta. 1986 Mar 7;870(1):76-81. doi: 10.1016/0167-4838(86)90010-5.
The radiation-inactivation size of membrane-bound acid beta-glucosidase in cultured skin fibroblasts of four normal individuals, five Gaucher type 1 (non-neuropathic), four Gaucher type 2 (acute neuropathic) and three Gaucher type 3 (sub-acute neuropathic) patients was determined using the radiation-inactivation method. The radiation-inactivation size of the enzyme in the control, Gaucher type 2 and Gaucher type 3 fibroblasts ranged from 94 000 to 128 800, and no statistical significant difference was found in the enzyme size between the normal and Gaucher cells nor among the Gaucher type 2 and type 3 cells. Contrary to the normal, Gaucher type 2 and Gaucher type 3 enzyme, the radiation-inactivation size of membrane-bound acid beta-glucosidase in all of the Gaucher type 1 fibroblasts tested is significantly higher, ranging from 158 400 to 235 300. The size of the control lysosomal enzyme, sphingomyelinase, also determined by the radiation-inactivation method in fibroblasts of normal individuals and patients with the three Gaucher subtypes, was between 70 000 and 74 500 and indistinguishable from each other. Since the molecular weight of acid beta-glucosidase subunit determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis was about 60 000 (Pentchev, P.G., Brady, R.O., Hibbert, S.P., Gal, A.E. and Shapiro, C. (1973) J. Biol. Chem. 248, 5256-5261), the above data suggest that: (i) the normal fibroblast enzyme, as well as the Gaucher type 2 and type 3 mutant enzyme, in the membrane-bound form, exists as a dimer; (ii) the underlying biochemical and genetic defect in non-neuropathic (type 1) and neuropathic (type 2 and type 3) Gaucher disease is very different from each other; and (iii) subunit interaction of the mutant enzyme may be present in Gaucher type 1 fibroblasts, resulting in the formation of a higher-molecular-weight aggregate.
采用辐射灭活法测定了4名正常个体、5名1型戈谢病（非神经病变型）患者、4名2型戈谢病（急性神经病变型）患者和3名3型戈谢病（亚急性神经病变型）患者培养的皮肤成纤维细胞中膜结合酸性β-葡萄糖苷酶的辐射灭活大小。对照、2型戈谢病和3型戈谢病成纤维细胞中该酶的辐射灭活大小在94000至128800之间，正常细胞与戈谢病细胞之间以及2型和3型戈谢病细胞之间的酶大小无统计学显著差异。与正常、2型戈谢病和3型戈谢病的酶相反，所有测试的1型戈谢病成纤维细胞中膜结合酸性β-葡萄糖苷酶的辐射灭活大小显著更高，在158400至235300之间。正常个体和三种戈谢病亚型患者的成纤维细胞中，通过辐射灭活法测定的对照溶酶体酶鞘磷脂酶的大小在70000至74500之间，彼此无差异。由于十二烷基硫酸钠聚丙烯酰胺凝胶电泳测定的酸性β-葡萄糖苷酶亚基的分子量约为60000（Pentchev, P.G., Brady, R.O., Hibbert, S.P., Gal, A.E.和Shapiro, C. (1973) J. Biol. Chem. 248, 5256 - 5261），上述数据表明：(i) 正常成纤维细胞酶以及2型和3型戈谢病突变酶以膜结合形式存在时为二聚体；(ii) 非神经病变型（1型）和神经病变型（2型和3型）戈谢病潜在的生化和遗传缺陷彼此非常不同；(iii) 1型戈谢病成纤维细胞中可能存在突变酶的亚基相互作用，导致形成更高分子量的聚集体。
