Maret A, Salvayre R, Samadi M, Douste-Blazy L
Enzyme. 1987;37(4):208-17. doi: 10.1159/000469264.
Lymphoid cell lines (LCL) from 3 adult patients with non-neuropathic Gaucher disease were established by Epstein-Barr virus (EBV) transformation and were investigated from the view of enzymology. Glucosylceramide-beta-glucosidase (GlcCer-beta-glucosidase) was present in soluble and particulate fraction of LCL from normal subjects and was deficient in type 1 Gaucher LCL; the deficiency of all molecular forms, shown by electrofocusing, indicates that they are coded by the same gene. The existence of two non-specific beta-glucosidases, one soluble (minor), the other membrane-bound (major), was demonstrated in leucocytes and LCL from normals; in Gaucher LCL, these were also present in a normal range. Characteristic properties of the non-specific membrane-bound beta-glucosidase were defined: lability at acidic pH and strong inhibitory effect by detergents. These properties allowed to discriminate it from the lysosomal GlcCer-beta-glucosidase and to define optimal assay conditions for determination of residual GlcCer-beta-glucosidase activity in Gaucher disease, using artificial substrate, without interference of non-specific membrane-bound beta-glucosidase. These results demonstrate that EBV-transformed LCL represent an accurate model system for enzymatic studies of Gaucher disease.
通过爱泼斯坦-巴尔病毒（EBV）转化建立了3例非神经病变型戈谢病成年患者的淋巴细胞系（LCL），并从酶学角度进行了研究。葡糖神经酰胺-β-葡萄糖苷酶（GlcCer-β-葡萄糖苷酶）存在于正常受试者LCL的可溶性和颗粒部分，而在1型戈谢病LCL中缺乏；电聚焦显示所有分子形式均缺乏，表明它们由同一基因编码。在正常人和LCL的白细胞中证实存在两种非特异性β-葡萄糖苷酶，一种是可溶性的（次要的），另一种是膜结合的（主要的）；在戈谢病LCL中，这些酶也在正常范围内。定义了非特异性膜结合β-葡萄糖苷酶的特性：在酸性pH下不稳定，受去污剂强烈抑制。这些特性使其能够与溶酶体GlcCer-β-葡萄糖苷酶区分开来，并确定了在不受到非特异性膜结合β-葡萄糖苷酶干扰的情况下，使用人工底物测定戈谢病中残余GlcCer-β-葡萄糖苷酶活性的最佳检测条件。这些结果表明，EBV转化的LCL代表了用于戈谢病酶学研究的精确模型系统。
