N-octyl-β-valienamine up-regulates activity of F213I mutant β-glucosidase in cultured cells:: a potential chemical chaperone therapy for Gaucher disease
N-octyl-β-valienamine up-regulates activity of F213I mutant β-glucosidase in cultured cells:: a potential chemical chaperone therapy for Gaucher disease
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DOI：
10.1016/j.bbadis.2004.03.007
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发表时间：
2004-08-04
影响因子：
6.2
通讯作者：
Suzuki, Y
中科院分区：
文献类型：
--
作者：
Lin, H;Sugimoto, Y;Suzuki, Y
戈谢病（GD）是最常见的鞘脂病形式，由β-葡萄糖苷酶（β-Glu）缺陷引起。碳水化合物模拟物N-辛基-β-维列胺（NOV）是β-Glu的抑制剂。当应用于培养的具有F213 I β-Glu突变的GD成纤维细胞时，NOV增加了突变酶的蛋白水平并上调了细胞酶活性。在F213 I纯合细胞中观察到NOV的最大作用，其中以30 μ M NOV处理4天导致酶活性增加约6倍，高达对照细胞中活性的约80%。NOV在具有其他β-Glu突变N370 S、L444 P、84 CG和RecN 10的细胞中无效。免疫荧光和细胞分级显示F213 I突变酶在经NOV处理的细胞的溶酶体中的定位。与此一致，NOV恢复了F213 I纯合细胞中C-14标记的葡萄糖神经酰胺的清除。F213 I突变体β-Glu在体外中性pH下迅速丧失其活性，并且这种pH依赖性活性丧失被NOV减弱。这些结果表明，NOV作为化学伴侣加速F213 I突变体β-Glu的转运和成熟，并且可能表明该化合物对GD的治疗价值。(C)2004 Elsevier B. V.保留所有权利。
Gaucher disease (GD) is the most common form of sphingolipidosis and is caused by a defect of beta-glucosidase (beta-Glu). A carbohydrate mimic N-octyl-beta-valienamine (NOV) is an inhibitor of beta-Glu. When applied to cultured GD fibroblasts with F213I beta-Glu mutation, NOV increased the protein level of the mutant enzyme and up-regulated cellular enzyme activity. The maximum effect of NOV was observed in F213I homozygous cells in which NOV treatment at 30 muM for 4 days caused a similar to 6-fold increase in the enzyme activity, up to similar to 80% of the activity in control cells. NOV was not effective in cells with other beta-Glu mutations, N370S, L444P, 84CG and RecNcil. Immunofluorescence and cell fractionation showed localization of the F213I mutant enzyme in the lysosomes of NOV-treated cells. Consistent with this, NOV restored clearance of C-14-labeled glucosylceramide in F213I homozygous cells. F213I mutant beta-Glu rapidly lost its activity at neutral pH in vitro and this pH-dependent loss of activity was attenuated by NOV These results suggest that NOV works as a chemical chaperone to accelerate transport and maturation of F213I mutant beta-Glu and may suggest a therapeutic value of this compound for GD. (C) 2004 Elsevier B.V. All rights reserved.