Gaucher Disease: Enzymatic and Molecular Studies
Gaucher Disease: Enzymatic and Molecular Studies
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戈谢病：酶学和分子研究
DOI：
--
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
F. Smith
中科院分区：
文献类型：
--
作者：
G. Grabowski;P. Graves;M. Grace;J. Bergmann;F. Smith
人酸性β-葡糖苷酶（β-Glc，D-葡糖基-N-酰基鞘氨醇葡糖水解酶，EC 3.2.1.45）是一种膜相关溶酶体酶，可切割其天然底物葡糖神经酰胺（GC）和合成β-葡糖苷的β糖苷键1，2。这种酶的活性缺陷导致戈谢病的亚型和变体3。正常成熟多肽的预测分子量为55，834 Da 4，5。正常的酶也含有7- 15%的碳水化合物6，7和广泛的糖苷重建发生在培养的成纤维细胞后8，9。除了信号序列剪切，没有其他的蛋白水解加工已被检测到与人类酶从几个来源7，9。一个催化必需的羧酸，Asp 443，已暂时分配附近的羧基末端的497个氨基酸的酶通过使用亲和标记。溴[3 H]conduritol B环氧化物，一种活性位点定向共价结合剂10。此外，动力学研究表明，活性位点内存在三个亚位点，对底物和/或底物的糖基头基和酰基或烷基部分具有特异性11，12。这些后者的子网站影响表观kcat值或结合常数，分别11。类似的研究还表明，活性位点内的相对亲水区域位于三个亚位点的交界处，并已提出对鞘氨醇的羟基和胺基具有特异性11。糖基结合位点含有用于结合葡萄糖C-2、C-3和C-4处适当配置的羟基的残基以及pKaapp=6.7的残基，其是形成二元（EI）复合物所必需的12。对这些亚位点的持续研究应提供对正常β-Glc的结构/功能相关性以及戈谢病亚型和变体中酶缺陷性质的深入了解。
Human acid β-glucosidase (β-Glc, D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) is a membrane associated lysosomal enzyme which cleaves the βglucosidic linkage of its natural substrate, glucosylceramide (GC), and synthetic β-glucosides1,2. The defective activity of this enzyme leads to the subtypes and variants of Gaucher disease3. The normal mature polypeptide has a predicted molecular mass of 55,834 Da4,5. The normal enzyme also contains 7- 15% carbohydrate6,7 and extensive glycosidic remodelling occurs post-translationally in cultured fibroblasts8,9. Except for signal sequence clipping, no other proteolytic processing has been detected with the human enzyme from several sources7,9. A catalytically essential carboxylate, Asp443, has been provisionally assigned near the carboxy terminus of this 497 amino acid enzyme by using the affinity label. Bromo[3H]conduritol B epoxide, an active sitedirected covalent inhibitor10. In addition, kinetic studies have suggested the presence of three subsites within the active site with specificities for the glycon head group and the acyl or alkyl moieties of substrates and/or inhibitors11,12. These latter subsites influence apparent kcat values or binding constants, respectively11. Similar studies have also suggested a relatively hydrophilic region within the active site which is located at the junction of the three subsites and has been proposed to have specificity for the hydroxyl and amine group of sphingosine11. The glycon binding site contains residues for binding of properly configured hydroxyl groups at C-2, C-3 and C-4 of glucose as well as a residue with pKaapp=6.7 which is necessary for the formation of binary (EI) complexes12. Continued study of these subsites should provide insight into structure/function correlations of normal β-Glc as well as into the nature of the enzymatic defects in the subtypes and variants of Gaucher disease.
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人溶酶体 β-葡萄糖苷酶：催化、糖苷配基和疏水结合位点的动力学特征。
影响因子：
3.9
作者：
Grabowski,GA;Gatt,S;Kruse,J;Desnick,RJ
通讯作者：
Desnick,RJ
人酸性 β-葡萄糖苷酶：在 N-烷基-脱氧野尻霉素-琼脂糖上亲和纯化正常胎盘和戈谢病脾酶。
DOI：
10.1159/000469336
发表时间：
1986
期刊：
Enzyme
影响因子：
--
作者：
Osiecki-Newman,KM;Fabbro,D;Dinur,T;Boas,S;Gatt,S;Legler,G;Desnick,RJ;Grabowski,GA
通讯作者：
Grabowski,GA
人葡萄糖脑苷脂酶 cDNA 的分子克隆和核苷酸序列。
DOI：
10.1073/pnas.82.21.7289
发表时间：
1985
影响因子：
11.1
作者：
Sorge,J;West,C;Westwood,B;Beutler,E
通讯作者：
Beutler,E
DOI：
--
发表时间：
1986-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
G. Grabowski;K. Osiecki-Newman;T. Dinur;D. Fabbro;G. Legler;S. Gatt;R. Desnick
通讯作者：
G. Grabowski;K. Osiecki-Newman;T. Dinur;D. Fabbro;G. Legler;S. Gatt;R. Desnick
正常成纤维细胞和 I 型、II 型和 III 型戈谢病患者的成纤维细胞中葡萄糖脑苷脂酶的加工。
DOI：
10.1073/pnas.83.19.7472
发表时间：
1986
影响因子：
11.1
作者：
Beutler,E;Kuhl,W
通讯作者：
Kuhl,W