Effects of carboxy-terminal modifications of proteinase 3 (PR3) on the recognition by PR3-ANCA
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Updated: Sep 27, 2026
Protein Misfolding Cyclic Amplification of Prions
Published on: November 7, 2012
Stephen A Capizzi1, Margaret A Viss, Amber M Hummel
1Thoracic Diseases Research Unit and Hematology Research Unit, Mayo Graduate School of Medicine, Mayo Clinic, Rochester, Minnesota 55905, USA.
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.
