Hodanová Katerina, Melková Zora, Horowitz Mia, Hrebícek Martin
Center for Integrated Genomics and Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University, Ke Karlovu 2, Praha 2, 128 08, Czech Republic.
Eur J Hum Genet. 2003 May;11(5):369-74. doi: 10.1038/sj.ejhg.5200974.
Gaucher disease, the most prevalent lysosomal storage disease, is characterised by a significant phenotypic variation caused by more than 150 mutations. In order to verify pathogenicity of mutations found in the Czech Gaucher population, the vaccinia expression system was used. The wild-type human beta-glucocerebrosidase cDNA and cDNAs carrying the mutations 72delC, 1326insT, 1263del55, S196P, N370S, L444P, G202E, D409H, T369M, L444P+V460V, and D409H+T369M were expressed in Gaucher fibroblast cell line (L444P/S107L), BSC40, and HeLa G cells. The enzymatic activity and immunological reactivity were analysed. Only beta-glucocerebrosidase-deficient fibroblasts were suitable for expression using plasmid transfection. The expressed beta-glucosidase activity of mutant glucocerebrosidases was in good correlation with the presumed severity of the mutations.
戈谢病是最常见的溶酶体贮积病，其特征是由150多种突变导致显著的表型变异。为了验证在捷克戈谢病患者群体中发现的突变的致病性，采用了痘苗表达系统。野生型人β-葡萄糖脑苷脂酶cDNA以及携带72delC、1326insT、1263del55、S196P、N370S、L444P、G202E、D409H、T369M、L444P + V460V和D409H + T369M突变的cDNA在戈谢病成纤维细胞系（L444P/S107L）、BSC40和HeLa G细胞中表达。对酶活性和免疫反应性进行了分析。只有β-葡萄糖脑苷脂酶缺陷的成纤维细胞适合使用质粒转染进行表达。突变型葡萄糖脑苷脂酶的表达β-葡萄糖苷酶活性与推测的突变严重程度密切相关。
