Preparation of asymmetric, cerebroside sulfate-containing phospholipid vesicles

Benvenuto Cestaro, Elvira Pistolesi, Norbert Hershkowitz, Shimon Gátt

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

A procedure is described which inserts asymmetrically cerebroside sulfate ('sulfatide') into the outer leaflet of bilayered phospholipid vesicles. Cerebroside sulfate is adsorbed onto a cellulose, filter-paper support and, when incubated with phosphatidylcholine vesicles is transferred to and inserted into the outer leaflet of these vesicles. This transfer occurs at, or above the transition temperature of the phospholipid and follows a similar pattern with small or larger ('fused') unilamellar vesicles. The transfer is linear with time for 1-2 h and is maximal after about 6 h, when the sulfatide content reaches about 6 mol% of the total quantity of phospholipid, corresponding to about 10 mol% of the phospholipids present in the outer layer. Initial rates of sulfatide transfer were somewhat increased when the vesicles contained a positively charged lipid (e.g. stearylamine) and decreased when this lipid was negatively charged (e.g. dicetyl phosphate) or hydrophobic (e.g. cholesterol). Divalent ions markedly inhibited sulfatide transfer and monovalent ions did so to a lesser degree. Once incorporated into the outer leaflet of the vesicle, the sulfatide could not be removed by washing with buffer, 1 M NaCl or 1 M urea.

Original languageEnglish
Pages (from-to)13-20
Number of pages8
JournalBiochimica et Biophysica Acta - Biomembranes
Volume685
Issue number1
DOIs
StatePublished - 8 Feb 1982
Externally publishedYes

Keywords

  • Asymmetry
  • Cerebroside sulfate
  • Dimyristoylphosphatidylcholine
  • Dipalmitoylphosphatidylcholine
  • Phospholipid vesicle
  • Sulfatide

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