Azide‐Tagged Sphingolipids: New Tools for Metabolic Flux Analysis

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.

[1]  K. Kabayama,et al.  Syntheses of lactosyl ceramide analogues carrying novel bifunctional BODIPY dyes directed towards the differential analysis of multiplexed glycosphingolipids by MS/MS using iTRAQ. , 2014, Chemical communications.

[2]  C. Thiele,et al.  Alkyne lipids as substrates for click chemistry-based in vitro enzymatic assays[S] , 2013, Journal of Lipid Research.

[3]  M. Fukata,et al.  2-Bromopalmitate analogues as activity-based probes to explore palmitoyl acyltransferases. , 2013, Journal of the American Chemical Society.

[4]  Carsten Schultz,et al.  Bifunktionalisierte Fettsäuren zur Visualisierung und Identifizierung von Protein‐Lipid‐Interaktionen in lebenden Zellen , 2013 .

[5]  A. Gavin,et al.  In vivo profiling and visualization of cellular protein-lipid interactions using bifunctional fatty acids. , 2013, Angewandte Chemie.

[6]  A. Shevchenko,et al.  Tracing fatty acid metabolism by click chemistry. , 2012, ACS chemical biology.

[7]  F. Goñi,et al.  In situ synthesis of fluorescent membrane lipids (ceramides) using click chemistry , 2012, Journal of chemical biology.

[8]  Y. Hannun,et al.  Ceramide synthases at the centre of sphingolipid metabolism and biology. , 2012, The Biochemical journal.

[9]  Xianlin Han,et al.  Multi-dimensional mass spectrometry-based shotgun lipidomics and novel strategies for lipidomic analyses. , 2012, Mass spectrometry reviews.

[10]  E. Lallana,et al.  Zuverlässige und effiziente Konjugation von Biomolekülen über Huisgen‐Azid‐Alkin‐Cycloadditionen , 2011 .

[11]  R. Riguera,et al.  Reliable and efficient procedures for the conjugation of biomolecules through Huisgen azide-alkyne cycloadditions. , 2011, Angewandte Chemie.

[12]  Stefan R Bornstein,et al.  Shotgun lipidomics on high resolution mass spectrometers. , 2011, Cold Spring Harbor perspectives in biology.

[13]  T. Kolter.  A view on sphingolipids and disease. , 2011, Chemistry and physics of lipids.

[14]  C. Bertozzi,et al.  From Mechanism to Mouse: A Tale of Two Bioorthogonal Reactions , 2011, Accounts of chemical research.

[15]  Carolyn R Bertozzi,et al.  Bringing chemistry to life , 2011, Nature Methods.

[16]  M. Debets,et al.  Bioconjugation with strained alkenes and alkynes. , 2011, Accounts of chemical research.

[17]  S. Takeoka,et al.  Multiplex analysis of sphingolipids using amine-reactive tags (iTRAQ) , 2011, Journal of Lipid Research.

[18]  A. Gomez-Muñoz,et al.  Control of metabolism and signaling of simple bioactive sphingolipids: Implications in disease. , 2010, Progress in lipid research.

[19]  G. Schmitz,et al.  A rapid and quantitative LC-MS/MS method to profile sphingolipids , 2010, Journal of Lipid Research.

[20]  Renliang Zhang,et al.  Quantification of ceramide species in biological samples by liquid chromatography electrospray ionization tandem mass spectrometry. , 2010, Analytical biochemistry.

[21]  Jeremy C. Allegood,et al.  Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers[S] , 2009, Journal of Lipid Research.

[22]  A. Llebaria,et al.  Synthesis and biological properties of Pachastrissamine (jaspine B) and diastereoisomeric jaspines. , 2009, Bioorganic & medicinal chemistry.

[23]  Yusuf A. Hannun,et al.  Principles of bioactive lipid signalling: lessons from sphingolipids , 2008, Nature Reviews Molecular Cell Biology.

[24]  J. Fernandez-Checa,et al.  Pharmacological modulation of sphingolipids and role in disease and cancer cell biology. , 2007, Mini reviews in medicinal chemistry.

[25]  T. Ullrich,et al.  Synthesis of borondipyrromethene (BODIPY)-labeled sphingosine derivatives by cross-metathesis reaction. , 2007, The Journal of organic chemistry.

[26]  M. Distefano,et al.  Selective Labeling of Proteins by Using Protein Farnesyltransferase , 2007, Chembiochem : a European journal of chemical biology.

[27]  A. Bielawska,et al.  Regulation of the sphingosine-recycling pathway for ceramide generation by oxidative stress, and its role in controlling c-Myc/Max function. , 2006, The Biochemical journal.

[28]  A. Merrill,et al.  Sphingolipidomics: high-throughput, structure-specific, and quantitative analysis of sphingolipids by liquid chromatography tandem mass spectrometry. , 2005, Methods.

[29]  R. Grubbs,et al.  A general model for selectivity in olefin cross metathesis. , 2003, Journal of the American Chemical Society.

[30]  F. Chevy,et al.  Coupled assay of sphingomyelin and ceramide molecular species by gas liquid chromatography. , 2002, Journal of lipid research.

[31]  M. Yano,et al.  Quantitative analysis of ceramide molecular species by high performance liquid chromatography. , 1998, Journal of Lipid Research.

[32]  M. Previati,et al.  Low nanogram range quantitation of diglycerides and ceramide by high-performance liquid chromatography. , 1996, Analytical biochemistry.

[33]  P. Herold,et al.  PB20 SYNTHESIS OF D-erythro- AND D-threo-SPHINGOSINE DERIVATIVES FROM L-SERINE , 1988 .

[34]  M. Iwamori,et al.  Analysis of ceramide and monohexaosyl glycolipid derivatives by high-performance liquid chromatography and its application to the determination of the molecular species in tissues. , 1987, Journal of chromatography.

[35]  J. Park,et al.  The synthesis and configurational stability of differentially protected .beta.-hydroxy-.alpha.-amino aldehydes , 1987 .

[36]  V. Armstrong,et al.  The reaction between thiols and 8-azidoadenosine derivatives. , 1976, Nucleic acids research.