[Dataset] A fluorogenic substrate for the detection of lipid amidases in intact cells

Lipid amidases of therapeutic relevance include acid ceramidase (AC), N-acylethanolamine-hydrolyzing acid amidase, and fatty acid amide hydrolase (FAAH). Although fluorogenic substrates have been developed for the three enzymes and high-throughput methods for screening have been reported, a platform...

Full description

Bibliographic Details
Authors: Casasampere, Mireia, Ung, Johnson, Iñáñez, Alejandro, Dufau, Carine, Tsuboi, Kazuhito, Casas, Josefina, Tan, Su-Fern, Feith, David J, Andrieu-Abadie, Nathalie, Segui, Bruno, Loughran, Thomas P., Abad, José Luis, Fabriàs, Gemma
Format: conjunto de datos
Publication Date:2024
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/383422
Online Access:http://hdl.handle.net/10261/383422
https://digital.csic.es/handle/10261/353089
Access Level:Open access
Keyword:N-palmitoylethanolamine
N-acylethanolamine acid amidase
Lipids
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
Description
Summary:Lipid amidases of therapeutic relevance include acid ceramidase (AC), N-acylethanolamine-hydrolyzing acid amidase, and fatty acid amide hydrolase (FAAH). Although fluorogenic substrates have been developed for the three enzymes and high-throughput methods for screening have been reported, a platform for the specific detection of these enzyme activities in intact cells is lacking. In this article, we report on the coumarinic 1-deoxydihydroceramide RBM1-151, a 1-deoxy derivative and vinilog of RBM14-C12, as a novel substrate of amidases. This compound is hydrolyzed by AC (appKm = 7.0 μM; appVmax = 99.3 nM/min), N-acylethanolamine-hydrolyzing acid amidase (appKm = 0.73 μM; appVmax = 0.24 nM/min), and FAAH (appKm = 3.6 μM; appVmax = 7.6 nM/min) but not by other ceramidases. We provide proof of concept that the use of RBM1-151 in combination with reported irreversible inhibitors of AC and FAAH allows the determination in parallel of the three amidase activities in single experiments in intact cells.

Similar Items