Topology of proteolipid protein in the myelin membrane of central nervous system. A study using antipeptide antibodies.
Peptides according to amino-acid sequences of the N- and C-terminus of lipophilin (proteolipid protein, PLP) (Gly1-Phe15 = 1; Thr261-Phe276 = 6) and of the other four hydrophilic domains (Glu37-Leu60 = 2; Arg97-Leu112 = 3; Gly119-Gly127 = 3A; Trp144-Tyr156 = 3B; Lys191-Ala203 = 4; Asn222-Phe232 = 5) have been synthesized by the solid-phase Fmoc method, linked covalently to keyhole limpet hemocyanin (KLH) and used as antigens. Monospecific antibodies against these antigens were isolated by affinity chromatography. Each antibody recognized its epitope in isolated partially delipidated PLP with the ELISA technique, western blot, thin sections of paraffin embedded rat brains and in the plasma membrane of appropriately fixed/permeabilized rat oligodendrocytes in culture. After fixation with formaldehyde antipeptide 3A antibody stained intact non-permeabilized cells. Therefore the epitope 3A must be located on the extracellular surface of the membrane. This is in full support of our previous biochemical results on the orientation of lipophilin in the myelin membrane.
Stoffel
W
W
Institut für Physiologische Chemie, Köln.
Subkowski
T
T
Jander
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D001923:Brain Chemistry; D002417:Cattle; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D008969:Molecular Sequence Data; D009186:Myelin Sheath / Q000032:analysis* / Q000276:immunology; D009457:Neuroglia / Q000648:ultrastructure; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology*; D011487:Protein Conformation; D011510:Proteolipids / Q000032:analysis* / Q000276:immunology; D051381:Rats
D000906:Antibodies; D000939:Epitopes; D010455:Peptides; D011510:Proteolipids