DIFFERENTIATION OF BETA-GLUCOCEREBROSIDASE FROM BETA-GLUCOSIDASE IN HUMAN TISSUES USING SODIUM TAUROCHOLATE
DIFFERENTIATION OF BETA-GLUCOCEREBROSIDASE FROM BETA-GLUCOSIDASE IN HUMAN TISSUES USING SODIUM TAUROCHOLATE
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DOI：
10.1016/0003-9861(76)90547-6
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发表时间：
1976-01-01
影响因子：
3.9
通讯作者：
GLEW, RH
中科院分区：
文献类型：
--
作者：
PETERS, SP;COYLE, P;GLEW, RH
人体组织含有至少2种能够从4-甲基伞形基-β-D-吡喃葡萄糖苷释放葡萄糖的酶，但这些酶中只有2种可以水解葡萄糖脑苷脂，并且在患有戈谢氏病的个体中缺乏。在人肝脏中，这些2β-葡萄糖苷酶在其亚细胞定位、离子交换柱上的色谱行为、底物特异性以及对牛磺胆酸钠和磷脂的抑制或激活的敏感性方面有所不同。当相对非特异性的人工β-葡萄糖苷底物4-甲基伞形基-β-D-吡喃葡萄糖苷在最适合葡萄糖脑苷脂水解的测定条件下使用时，其可有效测量相对葡萄糖脑苷脂：β-葡萄糖苷酶活性，并且可用于评估个体关于戈谢病的状态。这些测定条件需要 pH 值接近中性（pH 5.5-6.5）并且存在洗涤剂牛磺胆酸钠。在使用4-甲基伞形基-β-D-吡喃葡萄糖苷作为底物的测定中包含牛磺胆酸钠允许特异性测量葡萄糖脑苷脂：β-葡萄糖苷酶活性，因为牛磺胆酸钠抑制与戈谢病无关的非特异性β-葡萄糖苷酶并刺激相关的β-葡萄糖脑苷酶活性。
Human tissues contain at least 2 enzymes capable of releasing glucose from 4-methylumbelliferyl-.beta.-D-glucopyranoside, but only 2 of these enzymes can hydrolyze glucocerebroside and is deficient in individuals with Gaucher''s disease. In human liver, these 2 .beta.-glucosidases differ in terms of their subcellular localization, chromatographic behavior on ion-exchange columns, substrate specificity and sensitivity to inhibition or activation by sodium taurocholate and phospholipids. When the relatively nonspecific, artificial .beta.-glucoside substrate, 4-methylumbelliferyl-.beta.-D-glucopyranoside, is used under assay conditions optimal for glucocerebroside hydrolysis, it is effective in measuring relative glucocerebroside:.beta.-glucosidase activity and can be used to evaluate an individual''s status with respect to Gaucher''s disease. These conditions of assay require a pH near neutrality (pH 5.5-6.5) and the presence of the detergent sodium taurocholate. The inclusion of sodium taurocholate in assays using 4-methylumbelliferyl-.beta.-D-glucopyranoside as substrate permits the specific measurement of glucocerebroside:.beta.-glucosidase activity because sodium taurocholate inhibits the nonspecific .beta.-glucosidase not involved in Gaucher''s disease and stimulates the relevant .beta.-glucocerebrosidase activity.