Site-specific antibodies to rat myelin proteolipids directed against the C-terminal hexapeptide |
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Authors: | J L Nussbaum G Roussel E Wünsch P Jollès |
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Institution: | 1. Centre de Neurochimie du CNRS and Unité44 de l''INSERM, 5, rue Blaise Pascal, 67084 Strasbourg Cédex France;2. Max-Planck Institut für Biochemie, D-8033 Martinsried F.R.G.;3. Laboratoire des Protéines (UnitéCNRS No. 102 alliéeàl''INSERM), Universitéde Paris V, 45 rue des Saints-Pères, 75270 Paris Cédex 06 France |
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Abstract: | A site-specific antiserum against the rat myelin proteolipids was produced in rabbits by injection of a synthetic polypeptide composed of the C-terminal amino acids of the proteolipid sequence. The immunogenic hexapeptide H-Gly-Arg-Gly-Thr-Lys-Phe-OH was coupled to chicken egg-albumin with dimethylsuberimidate. Antibodies specific for this peptide reacted with the 2 myelin proteolipid protein bands after SDS polyacrylamide gel electrophoresis and electrophoretic transfer onto nitrocellulose. Immunocytochemical investigations with this anti-peptide antiserum showed that the Golgi complexes of the oligodendrocytes were highly labeled as noted previously with multivalent antibodies. Labeling of vesicles and discontinuous staining of the plasmalemma were also observed in the most actively myelinating oligodendrocytes. In contrast to previous results, the major dense line was free of staining; this may indicate that at this site the C-terminal hexapeptide is inaccessible to these antibodies and perhaps buried in the lipid bilayer, in disagreement with the proposed organization of the myelin proteolipid in the myelin membrane. |
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Keywords: | Myelin Oligodendrocyte Proteolipids Site-specific antibody |
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