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Characterization of the CD177 interaction with the ANCA antigen proteinase 3

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Item Type:Article
Title:Characterization of the CD177 interaction with the ANCA antigen proteinase 3
Creators Name:Jerke, U. and Marino, S.F. and Daumke, O. and Kettritz, R.
Abstract:Proteinase 3 is a serine protease found in neutrophil granules and on the extracellular neutrophil membrane (mPR3). mPR3 is a major antigen for anti-neutrophil cytoplasmic antibodies (PR3-ANCAs), autoantibodies causing fatal autoimmune diseases. In most individuals, a subpopulation of neutrophils also produce CD177, proposed to present additional PR3 on the surface, resulting in CD177(neg)/mPR3(low) and CD177(pos)/mPR3(high) neutrophil subsets. A positive correlation has been shown between mPR3 abundance, disease incidence, and clinical outcome. We present here a detailed investigation of the PR3:CD177 complex, verifying the interaction, demonstrating the effect of binding on PR3 proteolytic activity and explaining the accessibility of major PR3-ANCA epitopes. We observed high affinity PR3:CD177 complex formation by surface plasmon resonance. Using flow cytometry and a PR3-specific FRET assay, we found that CD177 binding reduced the proteolytic activity of PR3 in vitro using purified proteins, in neutrophil degranulation supernatants containing wtPR3 and directly on mPR3(high) neutrophils and PR3-loaded HEK cells. Finally, CD177(pos)/mPR3(high) neutrophils showed no migration advantage in vitro or in vivo when migrating from the blood into the oral cavity. We illuminate details of the PR3:CD177 interaction explaining mPR3 membrane orientation and proteolytic activity with relevance to ANCA activation of the distinct mPR3 neutrophil populations.
Keywords:Antineutrophil Cytoplasmic, Antibodies, Autoimmunity, Cell Degranulation, Cell Surface, Receptors, Endothelial Cells, Epitopes, GPI-Linked Proteins, Gene Expression Regulation, HEK293 Cells, Human Umbilical Vein Endothelial Cells, Isoantigens, Molecular, Models, Myeloblastin, Neutrophils, Primary Cell Culture, Protein Binding, Signal Transduction, Surface Plasmon Resonance
Source:Scientific Reports
ISSN:2045-2322
Publisher:Nature Publishing Group (U.K.)
Volume:7
Page Range:43328
Date:27 February 2017
Official Publication:https://doi.org/10.1038/srep43328
PubMed:View item in PubMed

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