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Título: | Activity of neutral and alkaline ceramidases on fluorogenic N -acylated coumarin-containing aminodiols |
Autor: | Casasampere, Mireia CSIC ORCID; Camacho, Luz CSIC ORCID; Casas, Josefina CSIC ORCID ; Cingolani, Francesca CSIC; Egido-Gabás, Meritxell CSIC; Abad, José Luis CSIC ORCID ; Bedia, Carmen CSIC ORCID; Fabriàs, Gemma CSIC ORCID | Palabras clave: | Ceramides Sphingolipids Acid ciramidase |
Fecha de publicación: | 1-oct-2015 | Editor: | American Society for Biochemistry and Molecular Biology | Citación: | Journal of Lipid Research 56 (10): 2019-2028 (2015) | Resumen: | Ceramidases catalyze the cleavage of ceramides into sphingosine and fatty acids. Previously, we reported on the use of the RBM14 fluorogenic ceramide analogs to determine acidic ceramidase activity. In this work, we investigated the activity of other amidohydrolases on RBM14 compounds. Both bacterial and human purified neutral ceramidases (NCs), as well as ectopically expressed mouse neutral ceramidase hydrolyzed RBM14 with different selectivity, depending on the N -acyl chain length. On the other hand, microsomes from alkaline ceramidase (ACER)3 knockdown cells were less competent at hydrolyzing RBM14C12, RBM12C14, and RBM14C16 than controls, while microsomes from ACER2 and ACER3 overexpressing cells showed no activity toward the RBM14 substrates. Conversely, N -acylethanolamine-hydrolyzing acid amidase (NAAA) overexpressing cells hydrolyzed RBM14C14 and RBM14C16 at acidic pH. Overall, NC, ACER3, and, to a lesser extent, NAAA hydrolyze fluorogenic RBM14 compounds. Although the selectivity of the substrates toward ceramidases can be modulated by the length of the N -acyl chain, none of them was specific for a particular enzyme. Despite the lack of specificity, these substrates should prove useful in library screening programs aimed at identifying potent and selective inhibitors for NC and ACER3. Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc. | Versión del editor: | http://dx.doi.org/10.1194/jlr.D061564 | URI: | http://hdl.handle.net/10261/178238 | DOI: | 10.1194/jlr.D061564 |
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Activity of neutral and alkaline ceramidases on fluorogenic N -acylated coumarin-containing aminodiols.pdf | 1,73 MB | Adobe PDF | Visualizar/Abrir |
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