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

FEBS Letters

Descamps-Latscha, Béatrice¹  Almeida, Roque P.³  Melchior, Maxine³  Gabay, Joëlle E.³  Nusbaum, Patrick²  Jamaleddine, Ghassan³  Witko-Sarsat, Véronique¹  Halbwachs-Mecarelli, Lise²  Lesavre, Philippe² 

[1] INSERM U25, Hôpital Necker, 161 rue de Sèvres, 75015 Paris, France;INSERM U90, Hôpital Necker, 161 rue de Sèvres, 75015 Paris, France;Beatrice and Samuel A. Seaver Laboratory, Division of Hematology-Oncology, Department of Medicine, Cornell University Medical College, New York, NY 10021, USA

关键词: ANCA;    Azurophil granule;    Baculovirus;    Proteinase 3;    Wegener;    AcMNPV;    Autographa californica mononuclear polyhedrosis virus;    AG;    azurophil granules;    ANCA;    anti-neutrophil cytoplasmic antibody;    cANCA;    cytoplasmic ANCA;    PR3;    proteinase 3;    Sf9;    Spodoptera frugiperda clone 9 cells;    WG;    Wegener's granulomatosis;   

DOI  :  10.1016/0014-5793(96)00152-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown