Molecular basis of lysosomal storage disease and the possibility of a new therapeutic approach

溶酶体贮积病的分子基础和新治疗方法的可能性

• 批准号: 10672178
• 负责人: OKUMIYA Toshika
• 金额: $ 1.98万
• 依托单位: Kochi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672178/
• 关键词: Iysosomal storage disease; Fabry disease; GMィイD21ィエD2-gangliosidosis; Morquio B disease; gene mutation; α-galactosidase; β-galactosidase; substrate analog; Morquio disease

(1) Tissue-specific posttranslational modification of human α-galactosidaseHeterogeneous gene expression and posttranslational glycosylation were examined in human α-galactosidase (α-Gal) transgenic mouse. The heart and liver of the mouse expressed a high amount of transcript as well as high α-Gal activity. Its gene products in the heart and kidney were sensitive to endoglycosidase H digestion, but those in the spleen and liver were largely resistant. Glycopeptidase F treatment confirmed an identical molecular mass for the peptide moiety of the enzyme. We conclude that heterogeneous molecular mass of the gene products is caused by different degree of posttranslational glycosylation in murine tissues.(2) Altered substrate specificity of mutant β-galactosidase in patients with Morquio B diseaseβ-Galactosidodosis is a lysosomal storage disease caused by a deficiency of acid β-galactosidase (β-Gal) consisting of two different clinical phenotypes, GM1-gangliosidosis and Morquio B disease. T … More he former has central nervous system lesions, whereas the latter has skeletal lesions with no central nervous system lesions. To clarify the molecular evidence to account for clinical difference between both diseases, we investigated substrate specificity of mutant β-Gal derived from patients with GM1-gangliosidosis and Morquio B disease using artificial substrate analogs. Mutant β-Gal (W237L/W237L, Y83H/R482C) from Morquio B disease exhibited relatively low affinity to Galβl-4Glc NAc , an analog of keratan sulfate, as compared with Galβ1-3GalNAc, an analog of GM1-gangliosid, but there was not such difference in both normal β-Gal and mutant β-Gal (R201C/R201C, 151T/151T) from GM1-gangliosidosis. Furthermore, Morquio B mutants showed considerable decrease in hydrolysis ratio for the analogs (Galβ1-4GlcNAc/Galβ1-3GalNAc), yet GM1-gangliosidosis mutants had similar hydrolysis ratio to normal β-Gal. We conclude that distinct clinical phenotypes between GM1-gangliosidosis and Moquio B disease are caused by altered substrate specificity of mutant P-Gal resulting from unique gene mutations in patients with Morquio B disease. Less

(1)人α-半乳糖苷酶的组织特异性翻译后修饰在人α-半乳糖苷酶(α-Gal)转基因小鼠中检查异质基因表达和翻译后糖基化。小鼠的心脏和肝脏表达大量转录物以及高α-Gal活性。其在心脏和肾脏中的基因产物对糖苷内切酶H的消化敏感，但在脾脏和肝脏中的基因产物很大程度上具有抵抗力。糖肽酶 F 处理证实了该酶的肽部分具有相同的分子量。我们得出结论，基因产物的异质分子量是由小鼠组织中不同程度的翻译后糖基化引起的。(2)Morquio B病患者突变型β-半乳糖苷酶底物特异性的改变β-半乳糖苷贮积症是一种由酸性β-半乳糖苷酶(β-Gal)缺乏引起的溶酶体贮积病，由两种不同的临床症状组成。 表型、GM1-神经节苷脂沉积症和 Morquio B 病。前者有中枢神经系统病变，后者有骨骼病变但无中枢神经系统病变。为了阐明解释这两种疾病之间临床差异的分子证据，我们使用人工底物类似物研究了源自 GM1-神经节苷脂贮积症和 Morquio B 病患者的突变 β-Gal 的底物特异性。与GM1-神经节苷脂类似物Galβ1-3GalNAc相比，来自Morquio B病的突变型β-Gal(W237L/W237L、Y83H/R482C)对硫酸角质素类似物Galβ1-4Glc NAc的亲和力相对较低，但正常β-Gal和突变型β-Gal均没有这种差异。 （R201C/R201C、151T/151T）来自 GM1-神经节苷脂贮积症。此外，Morquio B 突变体显示类似物 (Galβ1-4GlcNAc/Galβ1-3GalNAc) 的水解率显着降低，但 GM1-神经节苷脂沉积症突变体具有与正常 β-Gal 相似的水解率。我们得出的结论是，GM1-神经节苷脂贮积症和 Moquio B 病之间不同的临床表型是由 Morquio B 病患者的独特基因突变引起的突变 P-Gal 底物特异性改变引起的。较少的

项目成果

Satoshi Ishii, et al.: "α-Galactosidase transgenic mouse : Heterogeneous gene expression and posttranslational glycosylation in tissues"Glycoconjugate Journal. 15. 591-594 (1998)

Satoshi Ishii 等人：“α-半乳糖苷酶转基因小鼠：组织中的异质基因表达和翻译后糖基化”Glycoconjugate Journal 15. 591-594 (1998)。

Satoshi Ishii: "α-Galactosidase transgenic mouse:Heterogeneous gene expression and posttranslational glycosylation in tissues." Glycoconjugate Journal. 15. 591-594 (1998)

Satoshi Ishii：“α-半乳糖苷酶转基因小鼠：组织中的异质基因表达和翻译后糖基化。”糖缀合杂志。15. 591-594 (1998)

Satostli lshii: "α-Galactosidase transgenic mouse:Heterogeneous gene expression and posttranslational glycosylation in tissues"Glycoconjugate Journal. 15. 591-594 (1998)

Satostli lshii：“α-半乳糖苷酶转基因小鼠：组织中的异质基因表达和翻译后糖基化”《Glycoconjugate Journal》15. 591-594 (1998)。

Satoshi Ishii: "α-Galactosidase transgenic mouse:Hetergeneous gene expression and posttranslational glycosylation in tissues."Glycoconjugate Journal. 15. 591-594 (1998)

Satoshi Ishii：“α-半乳糖苷酶转基因小鼠：组织中的异质基因表达和翻译后糖基化。”糖缀合杂志 15. 591-594 (1998)