期刊
MOLECULAR BIOLOGY OF THE CELL
卷 28, 期 11, 页码 1507-1518出版社
AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E16-07-0536
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资金
- EMBO [ALTF 636-2013]
- Marie Skfodowska-Curie Intra-European Fellowships [MEMBRANE DYNAMICS-627088]
- Wolfson Foundation [18272]
- Medical Research Council [MC_UU_12010/Unit Programmes G0902418, MC_UU_12025]
- MRC/BBSRC/ESPRC [MR/K01577X/1]
- Wellcome Trust [107457/Z/15/Z, 104924/14/Z/14]
- Oxford-internal grants (John-Fell Fund)
- Oxford-internal grant (EPA Cephalosporin Fund)
- Scientific and Technological Research Council of Turkey [114Z192]
- EMBO Installation Grant [3024]
- Newton-Katip Celebi Fund
- BBSRC [BB/P026354/1] Funding Source: UKRI
- MRC [MC_UU_00008/9, MR/K01577X/1, MC_UU_12010/9] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/P026354/1, BB/L00433X/1] Funding Source: researchfish
- Medical Research Council [MR/K01577X/1, MC_UU_12010/9, MC_UU_00008/9] Funding Source: researchfish
Diffusion and interaction dynamics of molecules at the plasma membrane play an important role in cellular signaling and are suggested to be strongly associated with the actin cytoskeleton. Here we use superresolution 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, whereas transient nanodomain incorporation of ganglioside lipid GM1 is apparent in both the live-cell membrane and GPMVs. For GPI-APs, we detect two molecular pools in living cells; one pool shows high mobility with transient incorporation into nanodomains, and the other pool forms 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.
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