TY  - JOUR
A1  - Collenburg, Lena
A1  - Walter, Tim
A1  - Burgert, Anne
A1  - Mueller, Nora
A1  - Seibel, Juergen
A1  - Japtok, Lukasz
A1  - Kleuser, Burkhard
A1  - Sauer, Markus
A1  - Schneider-Schaulies, Sibylle
T1  - A Functionalized Sphingolipid Analogue for Studying Redistribution during Activation in Living T Cells
JF  - The journal of immunology
N2  - 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.
Y1  - 2016
U6  - https://doi.org/10.4049/jimmunol.1502447
SN  - 0022-1767
SN  - 1550-6606
VL  - 196
SP  - 3951
EP  - 3962
PB  - American Assoc. of Immunologists
CY  - Bethesda
ER  - 
TY  - GEN
A1  - Walter, Tim
A1  - Collenburg, Lena
A1  - Japtok, Lukasz
A1  - Kleuser, Burkhard
A1  - Schneider-Schaulies, Sibylle
A1  - Müller, Nora
A1  - Becam, Jerome
A1  - Schubert-Unkmeir, Alexandra
A1  - Kong, Ji Na
A1  - Bieberich, Erhard
A1  - Seibel, Jürgen
T1  - Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells
N2  - 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.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 324 
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-394960
ER  - 
TY  - JOUR
A1  - Walter, T.
A1  - Collenburg, Lena
A1  - Japtok, Lukasz
A1  - Kleuser, Burkhard
A1  - Schneider-Schaulies, Sibylle
A1  - Mueller, N.
A1  - Becam, Jerome
A1  - Schubert-Unkmeir, A.
A1  - Kong, J. N.
A1  - Bieberich, Erhard
A1  - Seibel, J.
T1  - Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells
JF  - Chemical communications
N2  - 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.
Y1  - 2016
U6  - https://doi.org/10.1039/c6cc02879a
SN  - 1359-7345
SN  - 1364-548X
VL  - 52
SP  - 8612
EP  - 8614
PB  - Royal Society of Chemistry
CY  - Cambridge
ER  -