Abstract
Non-vesicular lipid transport steps play a crucial role in lipid trafficking and potentially include spontaneous exchange. Since membrane contact facilitates this lipid transfer, it is most likely to occur at membrane contact sites (MCS). However, to date it is unknown whether closely attached biological membranes exchange lipids spontaneously. We have set up a system for studying the exchange of lipids at MCS formed between the endoplasmic reticulum (ER) and the plasma membrane. Contact sites were stably anchored and the lipids cholesterol and phosphatidylcholine (PC) were not capable of transferring spontaneously into the opposed bilayer. We conclude that physical contact between two associated biological membranes is not sufficient for transfer of the lipids PC and cholesterol.
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Abbreviations
- ER:
-
Endoplasmic reticulum
- PC:
-
Phosphatidylcholine
- PM:
-
Plasma membrane
- MCS:
-
Membrane contact site
- LTP:
-
Lipid transfer protein
- TIRF:
-
Total internal reflection fluorescence microscopy
- FLAP:
-
Fluorescence loss after photobleaching
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Acknowledgments
The authors would like to express their gratitude to Dr. Barbara Niemeyer for providing a plasmid encoding fluorescent protein-labelled STIM1 and Dr. Lars Kürschner for comments on the manuscript. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG, TRR83 to T.L.).
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Merklinger, E., Schloetel, JG., Spitta, L. et al. No Evidence for Spontaneous Lipid Transfer at ER–PM Membrane Contact Sites. J Membrane Biol 249, 41–56 (2016). https://doi.org/10.1007/s00232-015-9845-2
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DOI: https://doi.org/10.1007/s00232-015-9845-2