Synthesis and characterization of a new fluorogenic substrate for alpha-galactosidase.
Synthesis and characterization of a new fluorogenic substrate for alpha-galactosidase.
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DOI：
10.1007/s00216-009-2879-5
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发表时间：
2009-08
影响因子：
4.3
通讯作者：
Zheng, Wei
中科院分区：
文献类型：
--
作者：
Shi, Zhen-Dan;Motabar, Omid;Goldin, Ehud;Liu, Ke;Southall, Noel;Sidransky, Ellen;Austin, Christopher P.;Griffiths, Gary L.;Zheng, Wei
α-半乳糖苷酶A水解溶酶体中糖脂和糖蛋白的末端α-半乳糖基部分。α-半乳糖苷酶突变导致鞘糖脂、神经酰胺三己糖苷的溶酶体蓄积，从而导致法布里病。与突变酶蛋白结合并纠正其错误折叠和误折叠的小分子伴侣已成为法布里病的潜在疗法。我们合成了一种红色荧光底物试卤灵酰α-D-吡喃半乳糖苷，用于新的α-半乳糖苷酶的酶分析。由于试卤灵的pKa相对较低（约6），因此可在较低pH值下连续测定该含量，无需添加终止液。此外，该检测试剂盒发出红色荧光，与蓝色荧光相比，可显著降低化合物荧光和灰尘/棉绒造成的干扰。因此，这种新的红色荧光底物和由此产生的酶测定可用于高通量筛选，以确定法布里病的小分子伴侣。
Alpha-galactosidase A hydrolyzes the terminal alpha-galactosyl moieties from glycolipids and glycoproteins in lysosomes. Mutations in α-galactosidase cause lysosomal accumulation of the glycosphingolipid, globotriaosylceramide, which leads to Fabry disease. Small-molecule chaperones that bind to mutant enzyme proteins and correct their misfolding and mistrafficking have emerged as a potential therapy for Fabry disease. We have synthesized a red fluorogenic substrate, resorufinyl α-D-galactopyranoside, for a new α-galactosidase enzyme assay. This assay can be measured continuously at lower pH values, without the addition of a stop solution, due to the relatively low pKa of resorufin (~6). In addition, the assay emits red fluorescence, which can significantly reduce interferences due to compound fluorescence and dust/lint as compared to blue fluorescence. Therefore, this new red fluorogenic substrate and the resulting enzyme assay can be used in high-throughput screening to identify small-molecule chaperones for Fabry disease.
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影响因子：
3.9
作者：
YAGI, F;ECKHARDT, AE;GOLDSTEIN, IJ
通讯作者：
GOLDSTEIN, IJ
影响因子：
9.8
作者：
Wilcox, WR;Banikazemi, M;Germain, DP
通讯作者：
Germain, DP
影响因子：
4.1
作者：
Ishii, Satoshi;Chang, Hui-Hwa;Fan, Jian-Qiang
通讯作者：
Fan, Jian-Qiang
DOI：
10.1016/j.bbrc.2007.05.082
发表时间：
2007-07-20
影响因子：
3.1
作者：
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通讯作者：
Kaneski, Christine R.
影响因子：
2.9
作者：
TSOU, KC;SU, HCF
通讯作者：
SU, HCF