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Synthesis of Indoxyl-glycosides for Detection of Glycosidase Activities
Published on: May 27, 2015
Synthesis and characterization of a new fluorogenic substrate for alpha-galactosidase
Zhen-Dan Shi1, Omid Motabar, Ehud Goldin
1Imaging Probe Development Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-3708, USA.
Analytical and Bioanalytical Chemistry
|June 13, 2009
Summary
A new red fluorogenic assay for alpha-galactosidase was developed. This assay aids in discovering small-molecule chaperones for treating Fabry disease by enabling high-throughput screening.
Area of Science:
- Biochemistry
- Enzymology
- Lysosomal Storage Diseases
Background:
- Alpha-galactosidase A deficiency causes Fabry disease due to glycosphingolipid accumulation.
- Small-molecule chaperones are a potential therapeutic strategy for Fabry disease.
- Enzyme assays are crucial for identifying these therapeutic compounds.
Purpose of the Study:
- To develop a novel red fluorogenic substrate for alpha-galactosidase.
- To establish a continuous enzyme assay suitable for high-throughput screening.
- To facilitate the identification of small-molecule chaperones for Fabry disease.
Main Methods:
- Synthesis of resorufinyl alpha-D-galactopyranoside, a red fluorogenic substrate.
- Development of a continuous alpha-galactosidase enzyme assay.
- Characterization of the assay's performance at lower pH values and its fluorescence properties.
Main Results:
- The new assay utilizes a red fluorogenic substrate, resorufinyl alpha-D-galactopyranoside.
- The assay can be measured continuously at lower pH without a stop solution.
- Red fluorescence emission reduces interference from compound fluorescence and particulates.
Conclusions:
- The developed red fluorogenic assay is effective for alpha-galactosidase activity measurement.
- This assay is suitable for high-throughput screening to identify potential therapeutics for Fabry disease.
- The assay's properties offer advantages over traditional blue fluorescence assays.
