2024-03-28T11:55:34Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1782382022-03-24T13:48:52Zcom_10261_5063com_10261_5col_10261_5066
DIGITAL.CSIC
author
Casasampere, Mireia
author
Camacho, Luz
author
Casas, Josefina
author
Cingolani, Francesca
author
Egido-Gabás, Meritxell
author
Abad, José Luis
author
Bedia, Carmen
author
Fabriàs, Gemma
funder
Ministerio de Economía y Competitividad (España)
orcid
Fabriàs, Gemma [0000-0001-7162-3772]
orcid
Bedia, Carmen [0000-0003-4584-5843]
2019-03-21T10:11:52Z
2019-03-21T10:11:52Z
2015-10-01
Journal of Lipid Research 56 (10): 2019-2028 (2015)
http://hdl.handle.net/10261/178238
10.1194/jlr.D061564
http://dx.doi.org/10.13039/501100003329
26286360
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.
eng
openAccess
Ceramides
Sphingolipids
Acid ciramidase
Activity of neutral and alkaline ceramidases on fluorogenic N -acylated coumarin-containing aminodiols
artículo
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URL
https://digital.csic.es/bitstream/10261/178238/1/Activity%20of%20neutral%20and%20alkaline%20ceramidases%20on%20fluorogenic%20N%20-acylated%20coumarin-containing%20aminodiols.pdf
File
MD5
02ad111df89e7ef244caafc91619d0e8
1768324
application/pdf
Activity of neutral and alkaline ceramidases on fluorogenic N -acylated coumarin-containing aminodiols.pdf