Skip to Content
Merck

Azide-tagged sphingolipids: new tools for metabolic flux analysis.

Chembiochem : a European journal of chemical biology (2015-02-14)
María Garrido, José Luis Abad, Gemma Fabriàs, Josefina Casas, Antonio Delgado
ABSTRACT

Several diseases involve alterations in sphingolipid metabolism, so the development of tools for the analysis of sphingolipid metabolic fluxes is of interest. In this work, ω-azidosphingolipids 1-3 have been synthesized and tested as tracers in live cells. The synthesis starts from (S)-Garner's aldehyde and uses bromide or tosyloxy precursors for the introduction of the azido group into the sphingoid base. Studies in HGC-27 cells showed that probes 1-3 compete with the natural metabolites and are incorporated into sphingolipid pathways without affecting cell viability. The reactivity and bioorthogonality of the terminal azido group have been exploited by means of click reactions with different azadibenzocyclooctyne tags. This allows the mass spectrometric characterization of azidosphingolipidomes in pooled samples from different cell populations after independent treatments, providing proof of concept of the applicability of this technology in sphingolipid metabolic flux analysis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol-12C, 99.95 atom % 12C
Sigma-Aldrich
Aluminum oxide, nanopowder, 13 nm primary particle size (TEM), 99.8% trace metals basis
Sigma-Aldrich
Aluminum oxide, nanoparticles, <50 nm particle size (DLS), 20 wt. % in isopropanol
Sigma-Aldrich
Aluminum oxide, nanopowder, <50 nm particle size (TEM)
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Rhodium(III) nitrate solution, ~10 % (w/w) (Rh in >5 wt. % HNO3)
Sigma-Aldrich
Aluminum oxide, 99.997% trace metals basis
Supelco
Aluminum oxide, for the determination of hydrocarbons
Sigma-Aldrich
Aluminum oxide, nanowires, diam. × L 2-6 nm × 200-400 nm
Sigma-Aldrich
Aluminum oxide, single crystal substrate, <0001>
Sigma-Aldrich
Aluminum oxide, nanoparticles, 30-60 nm particle size (TEM), 20 wt. % in H2O
Sigma-Aldrich
Aluminum oxide, fused, powder, primarily α-phase, 100-200 mesh
Sigma-Aldrich
Aluminum oxide, powder, 99.99% trace metals basis
Sigma-Aldrich
Aluminum oxide, fused, powder, primarily α-phase, -325 mesh
Sigma-Aldrich
Aluminum oxide, Corundum, α-phase, -100 mesh
Sigma-Aldrich
Aluminum oxide, Brockmann I, standard grade, activated, acidic
Sigma-Aldrich
Aluminum oxide, Brockmann I, standard grade, activated, basic
Sigma-Aldrich
Aluminum oxide, activated, Brockmann I, standard grade, neutral
Sigma-Aldrich
Aluminum oxide, Type WN-6, Neutral, Activity Grade Super I
Sigma-Aldrich
Aluminum oxide, pore size 58 Å, ~150 mesh
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Methanol, NMR reference standard
Supelco
Methanol, analytical standard
Supelco
Aluminum oxide, activated, neutral, Brockmann Activity I
Sigma-Aldrich
Rhodium on alumina, extent of labeling: 5 wt. % loading, matrix alumina support, powder
Sigma-Aldrich
Rhodium, powder, 99.95% trace metals basis
Sigma-Aldrich
Aluminum oxide, mesoporous, MSU-X (wormhole), average pore size 3.8 nm
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Methanol, Absolute - Acetone free