Recombinant proteinase 3 (Wegener's antigen) expressed in Pichia pastoris is functionally active and is recognized by patient sera
- 1 November 1997
- journal article
- Published by Oxford University Press (OUP) in Clinical and Experimental Immunology
- Vol. 110 (2) , 257-264
- https://doi.org/10.1111/j.1365-2249.1997.tb08325.x
Abstract
SUMMARY: The open reading frame of human proteinase 3 (PR3) without the prepro-peptide was cloned and expressed in Escherichia coli (rcPR3) and in Pichia pastoris (rpPR3). The 6-histidine tagged rpPR3 was efficiently secreted into culture supernatant from which it could be purified by immobilized metal chelate chromatography. Purified rpPR3 migrated as a single 32-kD band on SDS-PAGE and harboured protease activity that could be inhibited with inhibitors specific for serine-proteases. By indirect antigen-capture ELISA using rpPR3, 60% of sera from patients with Wegener's granulomatosis bound to the recombinant product, although it was not recognized in ELISA with directly coated rpPR3.Keywords
This publication has 40 references indexed in Scilit:
- The Crystal Structure of PR3, a Neutrophil Serine Proteinase Antigen of Wegener's Granulomatosis AntibodiesJournal of Molecular Biology, 1996
- Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC‐1 cells is enzymatically active and recognized by c‐ANCAFEBS Letters, 1996
- Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti-neutrophil cytoplasmic autoantibodiesFEBS Letters, 1996
- Biosynthesis and Processing of Proteinase 3 in U937 CellsJournal of Biological Chemistry, 1996
- Inhibition of proteinase 3 by ANCA and its correlation with disease activity in Wegener's granulomatosisKidney International, 1995
- Secretion of human intracellular aspartic proteinase cathepsin E expressed in the methylotrophic yeast, Pichia pastoris and characterization of produced recombinant cathepsin EBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1994
- Cleavage and inactivation of human C1 inhibitor by the human leukocyte proteinase, proteinase 3European Journal of Immunology, 1993
- Relevance of classic anti-neutrophil cytoplasmic autoantibody (C-ANCA)-mediated inhibition of proteinase 3-α1-antitrypsin complexation to disease activity in Wegener's granulomatosisClinical and Experimental Immunology, 1993
- Interference of Wegener's granulomatosis autoantibodies with neutrophil Proteinase 3 activityClinical and Experimental Immunology, 1992
- Expression of dikaryon-specific mRNAs of Schizophyllum commune in relation to incompatibility genes, light, and fruitingExperimental Mycology, 1988