Journal article
Diffusion of lipids and GPI-anchored proteins in actin-free plasma membrane vesicles measured by STED-FCS.
- Abstract:
- Diffusion and interaction dynamics of molecules at the plasma membrane play an important role in cellular signalling, and they are suggested to be strongly associated with the actin cytoskeleton. Here, we utilise super-resolution STED microscopy combined with fluorescence correlation spectroscopy (STED-FCS) to access and compare the diffusion characteristics of fluorescent lipid analogues and GPI-anchored proteins (GPI-APs) in the live cell plasma membrane and in actin cytoskeleton-free cell-derived giant plasma membrane vesicles (GPMVs). Hindered diffusion of phospholipids and sphingolipids is abolished in the GPMVs while transient nanodomain incorporation of ganglioside lipid GM1 is apparent both in the live cell membrane and in GPMVs. For GPI-APs, we detect two molecular pools in living cells; one pool showing high mobility with transient incorporation into nanodomains, and the other pool forming immobile clusters, both of which disappear in GPMVs. Our data underline the crucial role of the actin cortex in maintaining hindered diffusion modes of many but not all of the membrane molecules, and highlight a powerful experimental approach to decipher specific influences on molecular plasma membrane dynamics.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 2.7MB, Terms of use)
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- Publisher copy:
- 10.1091/mbc.E16-07-0536
Authors
+ European Commission
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- Funding agency for:
- Sezgin, E
- Grant:
- Marie Skłodowska-Curie Intra-European Fellowships (MEMBRANE DYNAMICS-627088
+ EMBO
More from this funder
- Funding agency for:
- Sezgin, E
- Grant:
- Marie Skłodowska-Curie Intra-European Fellowships (MEMBRANE DYNAMICS-627088
- Grant:
- 3024
+ Scientific and Technological Research Council of Turkey
More from this funder
- Grant:
- TUBITAK, grant number 114Z192
- Publisher:
- American Society for Cell Biology
- Journal:
- Molecular Biology of the Cell More from this journal
- Volume:
- 28
- Issue:
- 11
- Pages:
- 1507-1518
- Publication date:
- 2017-04-12
- Acceptance date:
- 2017-04-04
- DOI:
- EISSN:
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1939-4586
- ISSN:
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1059-1524
- Language:
-
English
- Keywords:
- Pubs id:
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pubs:691167
- UUID:
-
uuid:1e49d975-0ca8-44e6-9923-f076adfd8b31
- Local pid:
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pubs:691167
- Source identifiers:
-
691167
- Deposit date:
-
2017-05-11
- ARK identifier:
Terms of use
- Copyright holder:
- Schneider et al
- Copyright date:
- 2017
- Notes:
- © 2017 Schneider et al. This article is distributed by The American Society for Cell Biology under license from the author(s). It is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
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