GPI (glycosylphosphatidylinositol)-anchored proteins are characteristic components of biochemically defined lipid rafts. Rafts may be involved in T-cell stimulation, but it is not clear whether molecules involved in TCR (T-cell receptor) signalling are partitioned to T-cell synapses through raft microdomains or through specific protein–protein interactions. We have used FRET (fluorescence resonance energy transfer) analysis to study the distribution of GPI-anchored fluorescent proteins in the plasma membrane of live cells. Multiple criteria suggested that FRET between different GPI-anchored fluorescent proteins in COS-7 or unstimulated Jurkat T-cells is generated by a random, unclustered distribution. Stimulation of TCR signalling in Jurkat T-cells by beads coated with antibodies against TCR subunits resulted in localized increases in fluorescence of raft markers. However, measurements of FRET and ratio imaging showed that there was no detectable clustering and no overall enrichment of raft markers in these regions.
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Conference Article|
October 26 2004
Distribution of lipid raft markers in live cells
O.O. Glebov;
O.O. Glebov
1
1MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, U.K.
1To whom correspondence should be addressed (email og@mrc-lmb.cam.ac.uk).
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B.J. Nichols
B.J. Nichols
1MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, U.K.
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Publisher: Portland Press Ltd
Received:
July 08 2004
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2004 The Biochemical Society
2004
Biochem Soc Trans (2004) 32 (5): 673–675.
Article history
Received:
July 08 2004
Citation
O.O. Glebov, B.J. Nichols; Distribution of lipid raft markers in live cells. Biochem Soc Trans 1 November 2004; 32 (5): 673–675. doi: https://doi.org/10.1042/BST0320673
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