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Selective Fluorogenic β-Glucocerebrosidase Substrates for Convenient Analysis of Enzyme Activity in Cell and Tissue Homogenates

Resource type
Date created
2020-02-28
Authors/Contributors
Abstract
Within mammals, there are often several functionally related glycoside hydrolases, which makes monitoring their activities problematic. This problem is particularly acute for the enzyme β-glucocerebrosidase (GCase), the malfunction of which is a key driver of Gaucher’s disease (GD) and a major risk factor for Parkinson’s disease (PD). Humans harbor two other functionally related β-glucosidases known as GBA2 and GBA3, and the currently used fluorogenic substrates are not selective, which has driven the use of complicated subtractive assays involving the use of detergents and inhibitors. Here we describe the preparation of fluorogenic substrates based on the widely used nonselective substrate resorufin β-d-glucopyranoside. Using recombinant enzymes, we show that these substrates are highly selective for GCase. We also demonstrate their value through the analysis of GCase activity in brain tissue homogenates from transgenic mice expressing mutant human GCase and patient fibroblasts expressing mutant GCase. This approach simplifies the analysis of cell and tissue homogenates and should facilitate the analysis of clinical and laboratory tissues and samples.
Description
The full text of this paper will be available in February, 2021 due to the embargo policies of Journal of the ACS Chemical Biology. Contact summit@sfu.ca to enquire if the full text of the accepted manuscript can be made available to you.
Identifier
DOI: 10.1021/acschembio.9b01044
Published as
Deen, M. C., Proceviat, C., Shan, X., Wu, L., Shen, D. L., Davies, G. J., & Vocadlo, D. J. (2020). Selective Fluorogenic β-Glucocerebrosidase Substrates for Convenient Analysis of Enzyme Activity in Cell and Tissue Homogenates. ACS Chemical Biology, 15(4), 824–829. https://doi.org/10.1021/acschembio.9b01044.
Publication details
Publication title
ACS Chemical Biology
Document title
Selective Fluorogenic β-Glucocerebrosidase Substrates for Convenient Analysis of Enzyme Activity in Cell and Tissue Homogenates
Date
2020
Volume
15
Issue
4
First page
824
Last page
829
Copyright statement
Copyright is held by the author(s).
Scholarly level
Peer reviewed?
Yes
Language
English

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