N-octyl-beta-valienamine up-regulates activity of F213I mutant beta-glucosidase in cultured cells: a potential chemical chaperone therapy for Gaucher disease |
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Authors: | Lin Hou Sugimoto Yuko Ohsaki Yuki Ninomiya Haruaki Oka Akira Taniguchi Miyako Ida Hiroyuki Eto Yoshikatsu Ogawa Seiichiro Matsuzaki Yuji Sawa Miwa Inoue Takehiko Higaki Katsumi Nanba Eiji Ohno Kousaku Suzuki Yoshiyuki |
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Affiliation: | Department of Neurobiology, Division of Child Neurology, Tottori University Faculty of Medicine, 86 Nishi-machi, Yonago 683-850, Japan. |
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Abstract: | 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 microM for 4 days caused a approximately 6-fold increase in the enzyme activity, up to approximately 80% of the activity in control cells. NOV was not effective in cells with other beta-Glu mutations, N370S, L444P, 84CG and RecNciI. 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 14C-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. |
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