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Title: Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs
Authors: Kinoshita, Masanao
Suzuki, Kenichi G.N.
Matsumori, Nobuaki
Takada, Misa
Ano, Hikaru
Morigaki, Kenichi
Abe, Mitsuhiro
Makino, Asami
Kobayashi, Toshihide
Hirosawa, Koichiro M.
Fujiwara, Takahiro K.
Kusumi, Akihiro
Murata, Michio
Author's alias: 木下, 祥尚
鈴木, 健一
松森, 信明
楠見, 明弘
村田, 道雄
Issue Date: 22-Mar-2017
Publisher: Rockefeller University Press
Journal title: The Journal of Cell Biology
Volume: 216
Issue: 4
Start page: 1183
End page: 1204
Abstract: Sphingomyelin (SM) has been proposed to form cholesterol-dependent raft domains and sphingolipid domains in the plasma membrane (PM). How SM contributes to the formation and function of these domains remains unknown, primarily because of the scarcity of suitable fluorescent SM analogs. We developed new fluorescent SM analogs by conjugating a hydrophilic fluorophore to the SM choline headgroup without eliminating its positive charge, via a hydrophilic nonaethylene glycol linker. The new analogs behaved similarly to the native SM in terms of their partitioning behaviors in artificial liquid order-disorder phase-separated membranes and detergent-resistant PM preparations. Single fluorescent molecule tracking in the live-cell PM revealed that they indirectly interact with each other in cholesterol- and sphingosine backbone{textendash}dependent manners, and that, for ~{}10{textendash}50 ms, they undergo transient colocalization-codiffusion with a glycosylphosphatidylinositol (GPI)-anchored protein, CD59 (in monomers, transient-dimer rafts, and clusters), in CD59-oligomer size{textendash}, cholesterol-, and GPI anchoring{textendash}dependent manners. These results suggest that SM continually and rapidly exchanges between CD59-associated raft domains and the bulk PM.
Description: 脂質の挙動をありのままに再現する蛍光プローブでラフトの形成機構を解明. 京都大学プレスリリース. 2017-03-28.
Rights: © 2017 Kinoshita et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http ://www .rupress .org /terms /). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https ://creativecommons .org /licenses /by -nc -sa /4 .0 /).
DOI(Published Version): 10.1083/jcb.201607086
PubMed ID: 28330937
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