Four-Color Single-Molecule Imaging with Engineered Tags Resolves the Molecular Architecture of Signaling Complexes in the Plasma Membrane

2021 ◽  
Author(s):  
Junel Sotolongo Bellón ◽  
Oliver Birkholz ◽  
Christian Paolo Richter ◽  
Florian Eull ◽  
Hella Kenneweg ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Single Molecule ◽  
Single Molecule Imaging ◽  
Molecular Architecture

scholarly journals High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

2020 ◽  
Vol 219 (12) ◽  
Author(s):  
Ikuko Koyama-Honda ◽  
Takahiro K. Fujiwara ◽  
Rinshi S. Kasai ◽  
Kenichi G.N. Suzuki ◽  
Eriko Kajikawa ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Plasma Membrane ◽  
Single Molecule ◽  
High Speed ◽  
Transmembrane Protein ◽  
Single Molecule Imaging ◽  
Lipid Interactions ◽  
Alkyl Chains ◽  
Outer Leaflet ◽  
Protein Moiety

Using single-molecule imaging with enhanced time resolutions down to 5 ms, we found that CD59 cluster rafts and GM1 cluster rafts were stably induced in the outer leaflet of the plasma membrane (PM), which triggered the activation of Lyn, H-Ras, and ERK and continually recruited Lyn and H-Ras right beneath them in the inner leaflet with dwell lifetimes <0.1 s. The detection was possible due to the enhanced time resolutions employed here. The recruitment depended on the PM cholesterol and saturated alkyl chains of Lyn and H-Ras, whereas it was blocked by the nonraftophilic transmembrane protein moiety and unsaturated alkyl chains linked to the inner-leaflet molecules. Because GM1 cluster rafts recruited Lyn and H-Ras as efficiently as CD59 cluster rafts, and because the protein moieties of Lyn and H-Ras were not required for the recruitment, we conclude that the transbilayer raft phases induced by the outer-leaflet stabilized rafts recruit lipid-anchored signaling molecules by lateral raft–lipid interactions and thus serve as a key signal transduction platform.

scholarly journals 1N0900 Single-molecule imaging of H-Ras and cRaf-1 on the plasma membrane of living cells

2002 ◽  
Vol 42 (supplement2) ◽  
pp. S80
Author(s):  
K. Hibino ◽  
Y. Sako ◽  
A. H. Iwane ◽  
T. Yanagida
Keyword(s):  
Plasma Membrane ◽  
Single Molecule ◽  
Living Cells ◽  
Single Molecule Imaging

scholarly journals Single-molecule imaging reveals dimerization/oligomerization of CXCR4 on plasma membrane closely related to its function

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Baosheng Ge ◽  
Jun Lao ◽  
Jiqiang Li ◽  
Yao Chen ◽  
Yanzhuo Song ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Single Molecule ◽  
Single Molecule Imaging

scholarly journals Single molecule imaging of activated small G protein Rac on the plasma membrane

2003 ◽  
Vol 43 (supplement) ◽  
pp. S101
Author(s):  
R. Nagai ◽  
T. Kobayashi ◽  
M. Murakami ◽  
T. Kawasaki ◽  
Y. Takeda ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
Single Molecule ◽  
Single Molecule Imaging ◽  
Small G Protein

scholarly journals Molecular Dynamics and Concentration in Raft and Boundary Domains in Actin-Depleted Plasma Membrane Vesicles as Revealed by Single-Molecule Imaging

2013 ◽  
Vol 104 (2) ◽  
pp. 191a
Author(s):  
Rahul Chadda ◽  
Takahiro K. Fujiwara ◽  
Ziya Kalay ◽  
Shusaku Shibutani ◽  
Kenichi G.N. Suzuki ◽  
...  
Keyword(s):  
Molecular Dynamics ◽  
Plasma Membrane ◽  
Single Molecule ◽  
Membrane Vesicles ◽  
Single Molecule Imaging ◽  
Plasma Membrane Vesicles

scholarly journals High-speed single-molecule imaging reveals signal transduction by induced transbilayer raft phases

2020 ◽  
Author(s):  
Ikuko Koyama-Honda ◽  
Takahiro K. Fujiwara ◽  
Rinshi S. Kasai ◽  
Kenichi G. N. Suzuki ◽  
Eriko Kajikawa ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Plasma Membrane ◽  
Single Molecule ◽  
High Speed ◽  
Transmembrane Protein ◽  
Single Molecule Imaging ◽  
Lipid Interactions ◽  
Alkyl Chains ◽  
Outer Leaflet ◽  
Protein Moiety

AbstractUsing single-molecule imaging with enhanced time resolutions down to 5 ms, we found that CD59-cluster rafts and GM1-cluster rafts stably induced in the outer leaflet of the plasma membrane (PM), which triggered the activation of Lyn, H-Ras, and ERK, continually recruited Lyn and H-Ras right beneath them in the inner leaflet, with dwell lifetimes of <0.1 s. The detection was possible due to the enhanced time resolutions employed here. The recruitment depended on the PM cholesterol and saturated alkyl chains of Lyn and H-Ras, whereas it was blocked by the non-raftophilic transmembrane protein moiety and unsaturated alkyl chains linked to the inner-leaflet molecules. Since GM1-cluster rafts recruited Lyn and H-Ras as efficiently as CD59-cluster rafts, and the protein moieties of Lyn and H-Ras were not required for the recruitment, we conclude that the transbilayer raft phases induced by the outer-leaflet stabilized rafts recruit lipid-anchored signaling molecules by lateral raft-lipid interactions, and thus serve as a key signal transduction platform.SummaryHigh-speed single-molecule imaging indicated that CD59-cluster rafts and GM1-cluster rafts stably induced in the plasma membrane outer leaflet generated nano-scale transbilayer raft phases, which continually and transiently recruited Lyn and H-Ras in the inner leaflet by cooperative raft-lipid interactions.

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