Phase separation of myelin sheath in Triton X-114 solution: Predominant localization of the 21.5-kDa isoform of myelin basic protein in the lipid raft-associated domain

Michihiro Uruse, Masahiro Yamamoto, Makoto Sugawa, Keiko Matsuura, Yurie Sato, Chika Seiwa, Kenji Watanabe, Sadakazu Aiso, Hiroaki Asou

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Myelin basic protein (MBP) isoforms in the myelin sheath are known to have distinct intracellular expression patterns, which are profoundly related to functional specificity. Determining the differential localization of MBP isoforms is therefore important for understanding their pathophysiological roles. In this study, we have developed a new method for phase separation of myelin. The non-ionic detergent Triton X-114 is used to solubilize myelin sheath which then undergoes phase separation to yield four fractions. The lipid raft-associated proteins and lipids in each fraction were analysed by immunoblotting and lipid analysis, respectively. The present method gives two lipid raft-enriched fractions, one of them was found to contain only lipid raft-associated galactocerebroside and cholesterol as the major lipids. The 21.5-kDa MBP isoforms (21.5 MBP), both unphosphorylated and phosphorylated, were exclusively contained in this fraction. Phosphorylated 21.5 MBP (21.5 pMBP) has been shown to specifically disappear from demyelinated loci. The present analytical method clearly indicated that disappearance of 21.5 pMBP corresponded to demyelination and its reappearance corresponded to prevention of demyelination. Demyelination was also associated with aging and was prevented by the myelin-protecting herbal medicine, Chinpi, a type of dried citrus peel.

Original languageEnglish
Pages (from-to)265-271
Number of pages7
JournalJournal of Biochemistry
Volume155
Issue number4
DOIs
Publication statusPublished - 2014 Jan 1
Externally publishedYes

Fingerprint

Myelin Basic Protein
Phase separation
Lipids
Protein Isoforms
Detergents
Nonidet P-40
Aging of materials
Cholesterol
Proteins

Keywords

  • 21.5-kDa isoform of myelin basic protein
  • demyelination
  • detergent extraction
  • lipid raft
  • remyelination
  • Triton X-114

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Phase separation of myelin sheath in Triton X-114 solution : Predominant localization of the 21.5-kDa isoform of myelin basic protein in the lipid raft-associated domain. / Uruse, Michihiro; Yamamoto, Masahiro; Sugawa, Makoto; Matsuura, Keiko; Sato, Yurie; Seiwa, Chika; Watanabe, Kenji; Aiso, Sadakazu; Asou, Hiroaki.

In: Journal of Biochemistry, Vol. 155, No. 4, 01.01.2014, p. 265-271.

Research output: Contribution to journalArticle

Uruse, Michihiro ; Yamamoto, Masahiro ; Sugawa, Makoto ; Matsuura, Keiko ; Sato, Yurie ; Seiwa, Chika ; Watanabe, Kenji ; Aiso, Sadakazu ; Asou, Hiroaki. / Phase separation of myelin sheath in Triton X-114 solution : Predominant localization of the 21.5-kDa isoform of myelin basic protein in the lipid raft-associated domain. In: Journal of Biochemistry. 2014 ; Vol. 155, No. 4. pp. 265-271.
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