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Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti‐neutrophil cytoplasmic autoantibodies
Author(s) -
Witko-Sarsat Véronique,
Halbwachs-Mecarelli Lise,
Almeida Roque P.,
Nusbaum Patrick,
Melchior Maxine,
Jamaleddine Ghassan,
Lesavre Philippe,
Descamps-Latscha Béatrice,
Gabay Joëlle E.
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00152-4
Subject(s) - proteinase 3 , recombinant dna , autoantibody , epitope , serine protease , chemistry , microbiology and biotechnology , antibody , immunology , biology , protease , biochemistry , enzyme , gene
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. Ollgosaccharides 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 of PR3, precluding its reactivity with sera from WG patients.