title: Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti-neutrophil cytoplasmic autoantibodies.
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.
authors.affiliation: INSERM U25, Hôpital Necker, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Witko-Sarsat
authors.forename: L
authors.initials: L
authors.lastname: Halbwachs-Mecarelli
authors.forename: R P
authors.initials: RP
authors.lastname: Almeida
authors.forename: P
authors.initials: P
authors.lastname: Nusbaum
authors.forename: M
authors.initials: M
authors.lastname: Melchior
authors.forename: G
authors.initials: G
authors.lastname: Jamaleddine
authors.forename: P
authors.initials: P
authors.lastname: Lesavre
authors.forename: B
authors.initials: B
authors.lastname: Descamps-Latscha
authors.forename: J E
authors.initials: JE
authors.lastname: Gabay
mesh_terms: D000581:Amidohydrolases / Q000494:pharmacology; D000818:Animals; D019268:Antibodies, Antineutrophil Cytoplasmic; D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology*; D002460:Cell Line; D000939:Epitopes / Q000032:analysis; D006031:Glycosylation; D014890:Granulomatosis with Polyangiitis / Q000201:enzymology / Q000276:immunology*; D006801:Humans; D007106:Immune Sera; D008565:Membrane Proteins; D008970:Molecular Weight; D053146:Myeloblastin; D017924:Nucleopolyhedroviruses / Q000235:genetics; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012697:Serine Endopeptidases / Q000737:chemistry / Q000276:immunology*; D012995:Solubility; D018411:Spodoptera; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D019268:Antibodies, Antineutrophil Cytoplasmic; D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D007106:Immune Sera; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D014415:Tunicamycin; D012697:Serine Endopeptidases; D053146:Myeloblastin; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase