Treatment of red blood cells with calcium and ionomycin causes activation of the lipid scramblase, a putative membrane protein catalysing flip-flop of (phospho)lipids. Various fluorescent 1-oleoyl-2-[6(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] caproyl (C6-NBD) analogues were tested for transbilayer movement across the plasma membrane of red blood cells. Among these phospholipid analogues were phosphatidylgalactose, phosphatidylmaltose and phosphatidylmaltotriose, which were obtained from C6-NBD-phosphatidylcholine by phospholipase D-catalysed transphosphatidylation. The inward movement after the onset of scrambling was monitored by extraction of the non-internalized probe with BSA. We demonstrate that both the amino group and the size of the headgroup determine the kinetics of lipid scrambling, and that lipids with a ceramide backbone migrate much more slowly than glycerophospholipids with the same headgroup.
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March 2002
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Research Article|
March 08 2002
Comparison between Ca2+-induced scrambling of various fluorescently labelled lipid analogues in red blood cells
David W. C. DEKKERS;
David W. C. DEKKERS
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Paul COMFURIUS;
Paul COMFURIUS
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Edouard M. BEVERS;
Edouard M. BEVERS
1
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
1To whom correspondence should be addressed (e-mail em.bevers@bioch.unimaas.nl).
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Robert F. A. ZWAAL
Robert F. A. ZWAAL
Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Publisher: Portland Press Ltd
Received:
July 27 2001
Revision Received:
November 14 2001
Accepted:
December 21 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 362 (3): 741–747.
Article history
Received:
July 27 2001
Revision Received:
November 14 2001
Accepted:
December 21 2001
Citation
David W. C. DEKKERS, Paul COMFURIUS, Edouard M. BEVERS, Robert F. A. ZWAAL; Comparison between Ca2+-induced scrambling of various fluorescently labelled lipid analogues in red blood cells. Biochem J 15 March 2002; 362 (3): 741–747. doi: https://doi.org/10.1042/bj3620741
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