A fluorogenic substrate for the detection of lipid amidases in intact cells.

ABSTRACT: 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 (^appK_m = 7.0 μM; ^appV_max = 99.3 nM/min), N-acylethanolamine-hydrolyzing acid amidase (^appK_m = 0.73 μM; ^appV_max = 0.24 nM/min), and FAAH

SUBMITTER: Casasampere M 

PROVIDER: S-EPMC10956054 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature