Developmental partitioning of myelin basic protein into membrane microdomains.
Specific membrane microdomains (including lipid rafts) exist in myelin but have not been fully characterized. Myelin basic protein (MBP) maintains the compactness of the myelin sheath and is highly posttranslationally modified. Thus, it has been suggested that MBP might also have other functions, e.g., in signal transduction. Here, the distribution of MBP and its modified forms was studied, spatially and temporally, by detailed characterization of membrane microdomains from developing and mature bovine myelin. Myelin membranes were extracted with three different detergents (Brij 96V, CHAPS, or Triton X-100) at 4 degrees C. The detergent-resistant membranes (DRMs), representing coalesced lipid rafts, were isolated as low-buoyant-density fractions on a sucrose density gradient. These myelin rafts were disrupted when cholesterol was depleted with methyl-beta-cyclodextrin. The use of CHAPS detergent led to enrichment of several myelin proteins, including phospho-Thr97-MBP, in the DRMs from mature myelin. Citrullinated and methylated MBP remained in "nonraft" microdomains. In contrast, the DRMs from early myelin were enriched in Golli-MBP, Fyn, Lyn, and CNP. The localization of various proteins in DRMs was further supported by the colocalization of these lipid raft components in cultured mouse oligodendrocytes. Thus, there is a developmental regulation of posttranslationally modified forms of MBP into specific membrane microdomains.
DeBruin
L S
LS
Department of Molecular and Cellular Biology and Biophysics Interdepartmental Group, University of Guelph, Guelph, Ontario, Canada.
Haines
J D
JD
Wellhauser
L A
LA
Radeva
G
G
Schonmann
V
V
Bienzle
D
D
Harauz
G
G
D000818:Animals; D001921:Brain / Q000196:embryology* / Q000254:growth & development / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D006651:Histocytochemistry; D021962:Membrane Microdomains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D009186:Myelin Sheath / Q000378:metabolism / Q000502:physiology*; D009836:Oligodendroglia / Q000378:metabolism; D011499:Protein Processing, Post-Translational
D004676:Myelin Basic Protein