Effects of carboxy-terminal modifications of proteinase 3 (PR3) on the recognition by PR3-ANCA

Stephen A. Capizzi, Margaret A. Viss, Amber M. Hummel, David N. Fass, Ulrich Specks

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Background. Autoantibodies directed against neutrophil proteinase 3 (PR3-ANCA) from patients with Wegener's granulomatosis and microscopic polyangiitis recognize conformational epitopes of PR3. During maturation of neutrophils, PR3 undergoes amino-terminal and carboxy-terminal processing. In contrast to amino-terminal processing, the effects of carboxy-terminal processing on recognition of PR3 by PR3-ANCA remain unknown. Carboxy-terminally modified or tagged recombinant PR3 (rPR3) molecules may be useful for the refinement of diagnostic assays and for the study of biological processes. Methods. This study was designed to determine whether 293 cells can be used to express specifically designed carboxy-terminal variants of rPR3, and to evaluate the effects of different carboxy-terminal modifications on the recognition by PR3-ANCA in the capture ELISA. Results. The rPR3-variants secreted into the media supernatants of transfected 293 cells escaped proteolytic processing. Furthermore, in contrast to the effects of amino-terminal pro-peptide deletion on PR3-ANCA binding, carboxy-terminal modifications (deletion and additions) did not significantly affect recognition by PR3-ANCA. Conclusions. This expression system is ideally suited for the expression of custom-designed carboxy-terminal rPR3 variants, and major conformational effects of carboxy-terminal modifications seem unlikely.

Original languageEnglish (US)
Pages (from-to)756-760
Number of pages5
JournalKidney international
Volume63
Issue number2
DOIs
StatePublished - 2003

Keywords

  • 293 cells
  • ANCA
  • Pauci-immune glomerulonephritis
  • Proteinase 3
  • Recombinant PR3
  • Small vessel vasculitis
  • Wegener's granulomatosis

ASJC Scopus subject areas

  • Nephrology

Fingerprint Dive into the research topics of 'Effects of carboxy-terminal modifications of proteinase 3 (PR3) on the recognition by PR3-ANCA'. Together they form a unique fingerprint.

Cite this