Identification and characterization of a B-cell determinant within the amphipathic domain (residues 178-238) of the myelin proteolipid protein

T. S. Stephens, M. Pakaski, M. B. Lees, N. T. Potter

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Pooled polyclonal rabbit anti-rat myelin and mouse anti-human proteolipid protein (PLP) antisera were screened against a panel of PLP synthetic peptides spanning residues 178-238 of the protein. Cross-reactivity against one determinant defined by PLP200-219 was particularly prominent in both the anti-myelin and anti-PLP antisera and was chosen for further study. Competitive inhibition studies, utilizing a panel of overlapping synthetic peptides, demonstrated that the C-terminal portion of PLP200-219, specifically residues comprising PLP200-217, was important for antibody recognition of this region. Immunohistochemical analyses with an affinity-purified rabbit anti-PLP200-219 antiserum demonstrated antibody cross-reactivity with PLP in both paraffin- and gelatin-embedded brain sections and immunocytochemical staining of mouse oligodendrocyte- enriched cultures demonstrated antibody binding with native PLP in situ. Staining of living non-permeabilized cells localized binding to the extracellular face of the myelin membrane. Collectively, these data argue for the presence of an immunodominant B-cell determinant defined by PLP residues 200-219. Furthermore, the structural conformation of this determinant in native PLP can be mimicked by the synthetic peptide, resulting in the generation of an antibody reagent that has considerable utility for immunohistochemical and immunocytochemical investigations of PLP expression and localization within the central nervous system myelin membrane.

Original languageEnglish
Pages (from-to)545-553
Number of pages9
JournalJournal of Neuroscience Research
Volume43
Issue number5
DOIs
Publication statusPublished - Mar 28 1996

Keywords

  • B-cell determinant
  • antibody
  • myelin
  • proteolipid protein
  • topography

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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