Activity of neutral and alkaline ceramidases on fluorogenic N-acylated coumarin-containing aminodiols
| العنوان: | Activity of neutral and alkaline ceramidases on fluorogenic N-acylated coumarin-containing aminodiols |
|---|---|
| المؤلفون: | Francesca Cingolani, Carmen Bedia, Ruijuan Xu, Mireia Casasampere, Yusuf A. Hannun, José Luis Abad, Meritxell Egido-Gabás, Kai Wang, Cungui Mao, Josefina Casas, Daniel Canals, Luz Camacho, Gemma Fabriàs |
| المساهمون: | Ministerio de Economía y Competitividad (España), Fabriàs, Gemma [0000-0001-7162-3772], Bedia, Carmen [0000-0003-4584-5843], Fabriàs, Gemma, Bedia, Carmen |
| المصدر: | Digital.CSIC. Repositorio Institucional del CSIC instname Journal of Lipid Research, Vol 56, Iss 10, Pp 2019-2028 (2015) |
| بيانات النشر: | The American Society for Biochemistry and Molecular Biology, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | Ceramide, Stereochemistry, Acylation, enzymology, Ceramidases, QD415-436, Alkaline Ceramidase, Ceramides, Biochemistry, Amidase, Substrate Specificity, lipids, chemistry.chemical_compound, Mice, Structure-Activity Relationship, umbelliferone, Endocrinology, Coumarins, Neutral Ceramidase, Methods, Animals, Humans, Fluorescent Dyes, mass spectrometry, chemistry.chemical_classification, Sphingolipids, ceramides, sphingolipids, Hydrolysis, Cell Biology, HCT116 Cells, Acid ciramidase, Enzyme, HEK293 Cells, chemistry, Ceramidase activity, Gene Knockdown Techniques, Microsome, Erratum, HeLa Cells |
| الوصف: | 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. This work was supported, in whole or in part, by the Spanish Ministry of Economy and Competitiveness (Grants SAF2011-22444 and CTQ2014-54743-R to G.F.), the Fundació la Marató de TV3 (Grants 112130 to J.C. and 112132), Agència de Gestió d’Ajuts Universitaris i de Recerca of Generalitat de Catalunya (Grant 2009SGR1072 to G.F.), National Cancer Institute (Grant NCI PO1 97132 to Y.A.H.), and National Institutes of Health (Grant R01CA163825 to C.M.). L.C. and F.C. were supported by predoctoral fellowships and C.B. was supported by postdoctoral contracts awarded by the Generalitat de Catalunya. |
| اللغة: | English |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7f588f5c644420e1808fb7870de940b9Test https://europepmc.org/articles/PMC4583077Test/ |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....7f588f5c644420e1808fb7870de940b9 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |