Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti-neutrophil cytoplasmic autoantibodies

FEBS Lett. 1996 Mar 11;382(1-2):130-6. doi: 10.1016/0014-5793(96)00152-4.

Abstract

Using the baculovirus/insect cells system, we have expressed a recombinant proteinase 3 (PR3) -- the neutrophil-derived serine protease autoantigen in Wegener's granulomatosis -- as a glycosylated intracellular and membrane-associated protein. Oligosaccharides accounted for the difference in molecular weights between recombinant (34 kDa) and neutrophil-PR3 (29 kDa). Whereas rabbit-anti-PR3 IgG recognized both recombinant and neutrophil-derived PR3, autoantibodies from Wegener patient sera recognized only neutrophil-derived PR3. Although oligosaccharides were not involved in PR3 epitope recognition, autoantibodies did not recognize the amino acid primary structure of recombinant PR3. Improper disulfide bond formation and/or lack of post-translational events in insect cells, may affect the conformation and/or lack of post-translational events in insect cells, may affect the conformation of PR3, precluding its reactivity with sera from WG patients.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amidohydrolases / pharmacology
  • Animals
  • Antibodies, Antineutrophil Cytoplasmic
  • Antigen-Antibody Reactions
  • Autoantibodies / immunology*
  • Autoantigens / chemistry
  • Autoantigens / immunology*
  • Cell Line
  • Epitopes / analysis
  • Glycosylation
  • Granulomatosis with Polyangiitis / enzymology
  • Granulomatosis with Polyangiitis / immunology*
  • Humans
  • Immune Sera
  • Membrane Proteins
  • Molecular Weight
  • Myeloblastin
  • Nucleopolyhedroviruses / genetics
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
  • Recombinant Fusion Proteins / biosynthesis
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / immunology*
  • Solubility
  • Spodoptera
  • Tunicamycin / pharmacology

Substances

  • Antibodies, Antineutrophil Cytoplasmic
  • Autoantibodies
  • Autoantigens
  • Epitopes
  • Immune Sera
  • Membrane Proteins
  • Recombinant Fusion Proteins
  • Tunicamycin
  • Serine Endopeptidases
  • Myeloblastin
  • Amidohydrolases
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase