Dokument-ID Dokumenttyp Verfasser/Autoren Herausgeber Haupttitel Abstract Auflage Verlagsort Verlag Erscheinungsjahr Seitenzahl Schriftenreihe Titel Schriftenreihe Bandzahl ISBN Quelle der Hochschulschrift Konferenzname Quelle:Titel Quelle:Jahrgang Quelle:Heftnummer Quelle:Erste Seite Quelle:Letzte Seite URN DOI Abteilungen OPUS4-45387 Wissenschaftlicher Artikel Collenburg, Lena; Walter, Tim; Burgert, Anne; Mueller, Nora; Seibel, Juergen; Japtok, Lukasz; Kleuser, Burkhard; Sauer, Markus; Schneider-Schaulies, Sibylle A Functionalized Sphingolipid Analogue for Studying Redistribution during Activation in Living T Cells Sphingolipids are major components of the plasma membrane. In particular, ceramide serves as an essential building hub for complex sphingolipids, but also as an organizer of membrane domains segregating receptors and signalosomes. Sphingomyelin breakdown as a result of sphingomyelinase activation after ligation of a variety of receptors is the predominant source of ceramides released at the plasma membrane. This especially applies to T lymphocytes where formation of ceramide-enriched membrane microdomains modulates TCR signaling. Because ceramide release and redistribution occur very rapidly in response to receptor ligation, novel tools to further study these processes in living T cells are urgently needed. To meet this demand, we synthesized nontoxic, azido-functionalized ceramides allowing for bio-orthogonal click-reactions to fluorescently label incorporated ceramides, and thus investigate formation of ceramide-enriched domains. Azido-functionalized C-6-ceramides were incorporated into and localized within plasma membrane microdomains and proximal vesicles in T cells. They segregated into clusters after TCR, and especially CD28 ligation, indicating efficient sorting into plasma membrane domains associated with T cell activation; this was abolished upon sphingomyelinase inhibition. Importantly, T cell activation was not abrogated upon incorporation of the compound, which was efficiently excluded from the immune synapse center as has previously been seen in Ab-based studies using fixed cells. Therefore, the functionalized ceramides are novel, highly potent tools to study the subcellular redistribution of ceramides in the course of T cell activation. Moreover, they will certainly also be generally applicable to studies addressing rapid stimulation-mediated ceramide release in living cells. Bethesda American Assoc. of Immunologists 2016 12 The journal of immunology 196 3951 3962 10.4049/jimmunol.1502447 Institut für Ernährungswissenschaft OPUS4-39496 misc Walter, Tim; Collenburg, Lena; Japtok, Lukasz; Kleuser, Burkhard; Schneider-Schaulies, Sibylle; Müller, Nora; Becam, Jerome; Schubert-Unkmeir, Alexandra; Kong, Ji Na; Bieberich, Erhard; Seibel, Jürgen Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells The synthesis and biological evaluation of azido-N-oleoyl serinol is reported. It mimicks biofunctional lipid ceramides and has shown to be capable of click reactions for cell membrane imaging in Jurkat and human brain microvascular endothelial cells. 2016 3 urn:nbn:de:kobv:517-opus4-394960 Institut für Ernährungswissenschaft OPUS4-45779 Wissenschaftlicher Artikel Walter, T.; Collenburg, Lena; Japtok, Lukasz; Kleuser, Burkhard; Schneider-Schaulies, Sibylle; Mueller, N.; Becam, Jerome; Schubert-Unkmeir, A.; Kong, J. N.; Bieberich, Erhard; Seibel, J. Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells The synthesis and biological evaluation of azido-N-oleoyl serinol is reported. It mimicks biofunctional lipid ceramides and has shown to be capable of click reactions for cell membrane imaging in Jurkat and human brain microvascular endothelial cells. Cambridge Royal Society of Chemistry 2016 3 Chemical communications 52 8612 8614 10.1039/c6cc02879a Institut für Ernährungswissenschaft