Phase separation of myelin proteins in triton X-114: differential behavior of myelin basic protein in purified myelin and in cultured oligodendrocytes.
Rabbit central (CNS) and peripheral nervous system (PNS) myelin, as well as nonmyelinating pig oligodendrocytes in culture, were extracted at 0-4 degrees C with the nonionic detergent Triton X-114. The solubilized proteins were partitioned into the detergent-rich and detergent-depleted (aqueous) phases that form upon heating to 37 degrees C. The proteolipid protein (PLP), myelin-associated glycoprotein (MAG), myelin oligodendrocyte glycoprotein (MOG) and P0 extracted from myelin were found exclusively in the detergent phase which is characteristic of the intrinsic membrane proteins. This was also the case for Wolfgram protein (WP), although this protein lacks transmembrane domains. A small fraction of the MAG and MOG extracted from oligodendrocytes partitioned into the aqueous phase, suggesting an altered conformation outside myelin or a different state of glycosylation. P2 and myelin basic protein (MBP) showed distinct patterns of behavior. P2 was found mainly in the aqueous phase giving strong support to its theoretically predicted conformation. Eighty-nine percent of the MBP extracted from CNS myelin and 81% of the pure MBP partitioned into the detergent phase. Surprisingly, most of the MBP extracted from the oligodendrocytes was recovered in the aqueous phase. We speculate that, in these cells, a hydrophilic protein might bind to the MBP in a specific manner, thereby preventing it from binding inappropriately to cellular components before its insertion into myelin.
Bürgisser
P
P
Service de Pédiatrie, CHUV, Lausanne, Suisse.
Matthieu
J M
JM
D000818:Animals; D002478:Cells, Cultured; D002498:Centrifugation; D003902:Detergents; D051379:Mice; D004676:Myelin Basic Protein / Q000032:analysis*; D018993:Myelin P0 Protein; D018996:Myelin P2 Protein; D009185:Myelin Proteins / Q000032:analysis*; D018991:Myelin Proteolipid Protein; D009186:Myelin Sheath / Q000032:analysis*; D018992:Myelin-Associated Glycoprotein; D009457:Neuroglia / Q000032:analysis*; D017830:Octoxynol; D009836:Oligodendroglia / Q000032:analysis*; D011092:Polyethylene Glycols; D011487:Protein Conformation; D011817:Rabbits; D011863:Radioimmunoassay; D012995:Solubility
D003902:Detergents; D004676:Myelin Basic Protein; D018993:Myelin P0 Protein; D018996:Myelin P2 Protein; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D018992:Myelin-Associated Glycoprotein; C028312:Wolfgram proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40