Multilamellar packing of myelin modeled by lipid-bound MBP.
Membrane compaction and adhesion at the major dense line (cytoplasmic apposition) of myelin, particularly in the central nervous system (CNS), is typically attributed to myelin basic protein (MBP). To explore the role of MBP in myelin membrane adhesion, we attempted to reconstitute the major dense line of myelin from purified lipid-bound MBP, which is a detergent-soluble form of MBP that retains the binding of all the myelin lipids. Removal of detergent by long-term dialysis yielded a precipitate, which, when analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and thin-layer chromatography, contained MBP that was still associated with myelin lipids, but in different proportions than in the native membrane. Comparison of lipid composition among isolated myelin, MBP-free myelin lipids, and lipid-bound MBP aggregates showed that the lipid-bound form of the protein was specifically enriched in phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, phosphatidylinositol, and phosphatidylserine. Electron microscopy and x-ray diffraction demonstrated that the lipid-MBP complexes formed multilayers having periods of 70-85 A, which correspond in width to individual myelin membranes. By contrast, the lipids alone assembled as multilayers having a period of approximately 40 A. Thus, the detergent-soluble form of MBP, which is bound to lipids, might serve as a simple model for the cytoplasmic apposition of myelin.
Riccio
P
P
Department of Biology, Defense and A.F. Biotechnology, University of Basilicata, Potenza, Italy.
Fasano
A
A
Borenshtein
N
N
Bleve-Zacheo
T
T
Kirschner
D A
DA
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008055:Lipids / Q000737:chemistry*; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000648:ultrastructure; D009186:Myelin Sheath / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D014961:X-Ray Diffraction
D008055:Lipids; D004676:Myelin Basic Protein