New Fluorogenic Probes for Neutral and Alkaline Ceramidases
March 2019
in “
Journal of lipid research
”
TLDR New probes were created to effectively measure specific enzymes involved in fat metabolism, which could help develop new drugs.
The document from June 2019 details the creation of new fluorogenic probes, RBM14 and RBM15, designed to measure the activity of neutral and alkaline ceramidases, which play a role in sphingolipid metabolism. The study found that the RBM14C24:1 probe was a highly selective substrate for human neutral ceramidase, while the RBM15C18:1 probe was the most effective for ACER1 and ACER2 ceramidases. These probes are important for developing high-throughput screening methods for ceramidase inhibitors with potential therapeutic applications. The study also provided kinetic parameters for the hydrolysis of substrates by neutral ceramidase, with KM values ranging from 8 to over 250 µM and Kcat/KM values from 15.4 to 546.5 M⁻¹s⁻¹, indicating efficient enzyme-substrate interactions. The effectiveness of the probes was confirmed through multiple experiments with triplicates, showing significant differences in umbelliferone release under various conditions and between different ceramidases (P < 0.05). The research was supported by the Spanish Ministry of Science, Innovation and Universities, with contributions from Alex García and Pedro Rayo.
