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J Biol Chem. 2016 May 13;291(20):10476-89. doi: 10.1074/jbc.M115.698639. Epub 2016 Mar 9.

Proteinase 3 Is a Phosphatidylserine-binding Protein That Affects the Production and Function of Microvesicles.

Author information

1
From the INSERM, U1016, Institut Cochin, 75014 Paris, France, CNRS-UMR8104, 75014 Paris, France, Université Paris Descartes, Sorbonne Paris Cité, 75006 Paris, France, Center of Excellence, Labex Inflamex, 75013 Paris, France.
2
Université Paris Descartes, Sorbonne Paris Cité, 75006 Paris, France, INSERM, U970, Paris Cardiovascular Research Center PARCC, 75015 Paris, France.
3
INSERM, UMR1098, Université Bourgogne Franche-Comté, Etablissement Français du Sang Bourgogne Franche-Comté, 25000 Besançon, France, Center of Excellence, Labex LipSTIC, 25000 Besançon, France.
4
Departments of Informatics and Molecular Biology, University of Bergen, 5008 Bergen, Norway.
5
Center of Excellence, Labex Inflamex, 75013 Paris, France, INSERM U1149/CNRS ERL8252, Université Paris-Diderot, 75018 Paris, France.
6
Université Grenoble Alpes, Institut de Biologie Structurale (IBS), 38044 Grenoble, France, CNRS, IBS, 38044 Grenoble, France, and Commissariat à l'Energie Atomique, IBS, 38000 Grenoble, France.
7
From the INSERM, U1016, Institut Cochin, 75014 Paris, France, CNRS-UMR8104, 75014 Paris, France, Université Paris Descartes, Sorbonne Paris Cité, 75006 Paris, France, Center of Excellence, Labex Inflamex, 75013 Paris, France, veronique.witko@inserm.fr.

Abstract

Proteinase 3 (PR3), the autoantigen in granulomatosis with polyangiitis, is expressed at the plasma membrane of resting neutrophils, and this membrane expression increases during both activation and apoptosis. Using surface plasmon resonance and protein-lipid overlay assays, this study demonstrates that PR3 is a phosphatidylserine-binding protein and this interaction is dependent on the hydrophobic patch responsible for membrane anchorage. Molecular simulations suggest that PR3 interacts with phosphatidylserine via a small number of amino acids, which engage in long lasting interactions with the lipid heads. As phosphatidylserine is a major component of microvesicles (MVs), this study also examined the consequences of this interaction on MV production and function. PR3-expressing cells produced significantly fewer MVs during both activation and apoptosis, and this reduction was dependent on the ability of PR3 to associate with the membrane as mutating the hydrophobic patch restored MV production. Functionally, activation-evoked MVs from PR3-expressing cells induced a significantly larger respiratory burst in human neutrophils compared with control MVs. Conversely, MVs generated during apoptosis inhibited the basal respiratory burst in human neutrophils, and those generated from PR3-expressing cells hampered this inhibition. Given that membrane expression of PR3 is increased in patients with granulomatosis with polyangiitis, MVs generated from neutrophils expressing membrane PR3 may potentiate oxidative damage of endothelial cells and promote the systemic inflammation observed in this disease.

KEYWORDS:

inflammation; microparticles; microvesicles; molecular modeling; neutrophil; proteinase; proteinase 3; serine protease; vasculitis

PMID:
26961880
PMCID:
PMC4865899
DOI:
10.1074/jbc.M115.698639
[Indexed for MEDLINE]
Free PMC Article

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