The first fluorogenic sensor for sphingosine-1-phosphate lyase activity in intact cells

Sanllehí, Pol; Casasampere, Mireia; Abad, José Luı́s; Fabriás, Gemma; López, Olga; Bujons, Jordi; Casas, Josefina; Delgado, Antonio

doi:10.1039/c7cc01659j

Cited by 14 publications

(12 citation statements)

References 14 publications

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“…However, no activity was observed after the initial assays (results not shown), which was attributed to the inability of the probe to cross the cell membrane. Encapsulation into liposomes, similarly as reported by us for a related probe (16), was not feasible due to the low solubility of RBM14C24:1 in the required solvent system. Gratifyingly, the use of methyl-β-cyclodextrin (MBCD), a well-know system used to increase the solubility of non-polar substances, such as fatty acids, lipids, vitamins and cholesterol in several cell culture applications (19) proved efficient to allow the use of RBM14C24:1 as NC substrate in intact cells ( Figure 5B).…”

Section: Selectivity Of Probe Rbm14c24:1 For Ncmentioning

confidence: 83%

“…Probes RBM14 were obtained by N-acylation of the free base with the required carboxylic acids, following previously reported protocols (12,13). Probes RBM15 were inspired in our "second generation" probes for S1P lyase, in which the installation of a vinyl unit between the amino diol moiety and the umbelliferone reporter led to a significant increase of affinity, probably due to the closer similarity of the vinylogated probe with the natural substrate (16).…”

Section: Design and Synthesis Of The Probesmentioning

confidence: 99%

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New fluorogenic probes for neutral and alkaline ceramidases

Casasampere

Bielsa

Riba

et al. 2019

Journal of Lipid Research

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New fluorogenic ceramidase substrates derived from the N-acyl modification of our previously reported probes (RBM14) are reported. While none of the new probes were superior to the known RBM14C12 as acid ceramidase substrates, the corresponding nervonic acid amide (RBM14C24:1) is an efficient and selective substrate for the recombinant human neutral ceramidase, both in cell lysates and in intact cells. A second generation of substrates, incorporating the natural 2-(N-acylamino)-1,3-diol-4-ene framework (compounds RBM15) is also reported. Among them, the corresponding fatty acyl amides with an unsaturated N-acyl chain can be used as substrates to determine alkaline ceramidase (ACER)1 and ACER2 activities. In particular, compound RBM15C18:1 has emerged as the best fluorogenic probe reported so far to measure ACER1 and ACER2 activities in a 96-well plate format.

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Section: Selectivity Of Probe Rbm14c24:1 For Ncmentioning

confidence: 83%

Section: Design and Synthesis Of The Probesmentioning

confidence: 99%

New fluorogenic probes for neutral and alkaline ceramidases

Casasampere

Bielsa

Riba

et al. 2019

Journal of Lipid Research

Self Cite

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“…The anti -isomer 4 was then subjected to olefin cross-coupling metathesis with 1-tetradecene using Grubbs 2nd generation cat. under standard conditions to provide the protected 1-deoxy sphingoid base 5 with the desired E -configuration as the major product (71% yield) [ 36 , 37 , 38 , 39 , 40 ].…”

Section: Resultsmentioning

confidence: 99%

Stereoselective Synthesis of Novel Sphingoid Bases Utilized for Exploring the Secrets of Sphinx

Saied

2021

IJMS

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Sphingolipids are ubiquitous in eukaryotic plasma membranes and play major roles in human and animal physiology and disease. This class of lipids is usually defined as being derivatives of sphingosine, a long-chain 1,3-dihydroxy-2-amino alcohol. Various pathological conditions such as diabetes or neuropathy have been associated with changes in the sphingolipidome and an increased biosynthesis of structurally altered non-canonical sphingolipid derivatives. These unusual or non-canonical sphingolipids hold great promise as potential diagnostic markers. However, due to their low concentrations and the unavailability of suitable standards, the research to explore the secret of this class of ‘Sphinx’ lipids is ultimately hampered. Therefore, the development of efficient and facile syntheses of standard compounds is a key endeavor. Here, we present various chemical approaches for stereoselective synthesis and in-depth chemical characterization of a set of novel sphingoid bases which were recently utilized as valuable tools to explore the metabolism and biophysical properties of sphingolipids, but also to develop efficient analytical methods for their detection and quantification.

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“…Similar approaches have since then been used to synthesize SL containing modified LCB. [24][25][26] Moreover, 1-deoxysphingosine derivatives have been synthesized, starting from L-alanine, instead of serine. [27][28][29] Here, the weinreb amide of L-alanine 9 was reacted with allyl magnesium bromide to form the ketone 10 (scheme 2).…”

Section: Resultsmentioning

confidence: 99%