TY  - JOUR
A1  - Fink, Julian
A1  - Schumacher, Fabian
A1  - Schlegel, Jan
A1  - Stenzel, Philipp
A1  - Wigger, Dominik
A1  - Sauer, Markus
A1  - Kleuser, Burkhard
A1  - Seibel, Jürgen
T1  - Azidosphinganine enables metabolic labeling and detection of sphingolipid de novo synthesis
T2  - Organic & biomolecular chemistry : an international journal of synthetic, physical and biomolecular organic chemistry
N2  - Here were report the combination of biocompatible click chemistry of omega-azidosphinganine with fluorescence microscopy and mass spectrometry as a powerful tool to elaborate the sphingolipid metabolism. The azide probe was efficiently synthesized over 13 steps starting from l-serine in an overall yield of 20% and was used for live-cell fluorescence imaging of the endoplasmic reticulum in living cells by bioorthogonal click reaction with a DBCO-labeled fluorophore revealing that the incorporated analogue is mainly localized in the endoplasmic membrane like the endogenous species. A LC-MS(/MS)-based microsomal in vitro assay confirmed that omega-azidosphinganine mimics the natural species enabling the identification and analysis of metabolic breakdown products of sphinganine as a key starting intermediate in the complex sphingolipid biosynthetic pathways. Furthermore, the sphinganine-fluorophore conjugate after click reaction was enzymatically tolerated to form its dihydroceramide and ceramide metabolites. Thus, omega-azidosphinganine represents a useful biofunctional tool for metabolic investigations both by in vivo fluorescence imaging of the sphingolipid subcellular localization in the ER and by in vitro high-resolution mass spectrometry analysis. This should reveal novel insights of the molecular mechanisms sphingolipids and their processing enzymes have e.g. in infection.
Y1  - 2021
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/62845
SN  - 1477-0520
SN  - 1477-0539
VL  - 19
IS  - 10
SP  - 2203
EP  - 2212
PB  - Royal Society of Chemistry
CY  - Cambridge
ER  -