TY - JOUR
T1 - Bifunctional Sphingosine for Cell-Based Analysis of Protein-Sphingolipid Interactions
AU - Haberkant, Per
AU - Stein, Frank
AU - Höglinger, Doris
AU - Gerl, Mathias J.
AU - Brügger, Britta
AU - Van Veldhoven, Paul P.
AU - Krijgsveld, Jeroen
AU - Gavin, Anne Claude
AU - Schultz, Carsten
PY - 2016/1/15
Y1 - 2016/1/15
N2 - 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.
AB - 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.
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U2 - 10.1021/acschembio.5b00810
DO - 10.1021/acschembio.5b00810
M3 - Article
C2 - 26555438
AN - SCOPUS:84955060006
VL - 11
SP - 222
EP - 230
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 1
ER -