Bifunctional sphingosine for cell-based analysis of protein-sphingolipid interactions

Haberkant, Per; Stein, Frank; Höglinger, Doris; Gerl, Mathias J; Brügger, Britta; Van Veldhoven, Paul P; Krijgsveld, Jeroen; Gavin Perrin, Anne-Claude; Schultz, Carsten

doi:10.1021/acschembio.5b00810

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Title		Bifunctional sphingosine for cell-based analysis of protein-sphingolipid interactions
Authors		Haberkant, Per Stein, Frank Höglinger, Doris Gerl, Mathias J Brügger, Britta Van Veldhoven, Paul P Krijgsveld, Jeroen Gavin Perrin, Anne-Claude Schultz, Carsten show all authors [1 - 9]
Published in		ACS Chemical Biology. 2016, vol. 11, no. 1, p. 222-230
Abstract		Sphingolipids are essential structural components of cellular membranes and are crucial regulators of cellular processes. While current high-throughput approaches allow for the systematic mapping of interactions of soluble proteins with their lipid-binding partners, photo-cross-linking is the only technique that enables for the proteome-wide mapping of integral membrane proteins with their direct lipid environment. Here, we report the synthesis of a photoactivatable and clickable analog of sphingosine (pacSph). When administered to sphingosine-1-phosphate lyase deficient cells, pacSph allows its metabolic fate and the subcellular flux of de novo synthesized sphingolipids to be followed in a time-resolved manner. The chemoproteomic profiling yielded over 180 novel sphingolipid-binding proteins, of which we validated a number, demonstrating the unique value of this technique as a discovery tool. This work provides an important resource for the understanding of the global cellular interplay between sphingolipids and their interacting proteins.
Keywords		Humans — Molecular Structure — Protein Array Analysis — Protein Binding — Proteomics/methods — Reproducibility of Results — Sphingolipids/metabolism — Sphingosine/analogs & derivatives/chemical synthesis/chemistry
Identifiers		DOI: 10.1021/acschembio.5b00810 PMID: 26555438
Full text		Article (Published version) (5.8 MB) - Limited access to UNIGE
Research group		Métabolisme des lipides (1001)
Citation (ISO format)		HABERKANT, Per et al. Bifunctional sphingosine for cell-based analysis of protein-sphingolipid interactions. In: ACS Chemical Biology, 2016, vol. 11, n° 1, p. 222-230. doi: 10.1021/acschembio.5b00810 https://archive-ouverte.unige.ch/unige:124754