Synthesis of alpha-D-Glcp-(1 -> 2)-alpha-D-Glcp-(1->3)-alpha-D-Glcp-O-(CH2)(8)COOCH3 for use in the assay of alpha-glucosidase I activity

被引：18
|
作者：
Scaman, CH
Hindsgaul, O
Palcic, MM
Srivastava, OP
机构：
[1] UNIV ALBERTA,DEPT CHEM,EDMONTON,AB T6G 2G2,CANADA
[2] ALBERTA RES COUNCIL,CARBOHYDRATE RES PROGRAM,EDMONTON,AB T6H 5X2,CANADA
来源：
CARBOHYDRATE RESEARCH | 1996年 / 296卷
基金：
加拿大自然科学与工程研究理事会;
关键词：
alpha-glucosidase I; fluorescence-labelled oligosaccharide; tetramethylrhodamine; glycosidase assay;
D O I：
10.1016/S0008-6215(96)00242-X
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The chemical synthesis of alpha-D-Glcp-(1 --> 2)-alpha-D-Glc p-(1 --> 3)-alpha-D-Glcp-O-(CH2)(8)COOCH3 (9), a substrate specific for cr-glucosidase I, is reported. This enzyme removes the terminal alpha-D-Glcp unit to produce alpha-D-Glcp-(1 --> 3)-alpha-D-Glcp-O-(CH2)(8)COOCH3 (10). This is the first synthetic substrate described for glucosidase I that allows kinetic evaluation of substrates and inhibitors of this enzyme. Tetramethylrhodamine was coupled to 9 through an ethylenediamine linker to produce a brilliant red derivative. Addition of this fluorescent dye did not affect enzyme binding to the substrate, as determined by a comparison of the K-m value (1.3 mM). The fluorescent label allows visual detection of 2-3 pmol of product by TLC. (C) 1996 Elsevier Science Ltd.
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页码：203 / 213
页数：11
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