A Monoclonal Antibody (MCPR3-7) Interfering with the Activity of Proteinase 3 by an Allosteric Mechanism

Hinkofer, Lisa C.; Seidel, Susanne A. I.; Korkmaz, Brice; Silva, Francisco; Hummel, Amber M.; Braun, Dieter; Jenne, Dieter E.; Specks, Ulrich

doi:10.1074/jbc.m113.495770

Cited by 20 publications

(26 citation statements)

References 37 publications

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“…The PR3 hydrophobic patch should not impair access of low M r phosphonate inhibitors to the PR3 active site cleft. The weak PR3 m activity and its resistance to inhibition might be due to its conformation being distorted; we recently used the monoclonal Ab MCPR3-7 to show that PR3 is in equilibrium between a highly favored active conformation and an inactive conformation (46). This antibody inhibits PR3 activity by an allosteric mechanism and competes with the neutrophil membrane for the binding of PR3.…”

Section: Discussionmentioning

confidence: 99%

New Selective Peptidyl Di(chlorophenyl) Phosphonate Esters for Visualizing and Blocking Neutrophil Proteinase 3 in Human Diseases

Guarino

Łęgowska

Epinette

et al. 2014

Journal of Biological Chemistry

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Background: Proteinase 3 activity is poorly controlled by physiological inhibitors, and its biological function is not well understood. Results:We have designed irreversible phosphonate inhibitors based on structural differences between proteinase 3 and elastase. Conclusion: They selectively inhibit proteinase 3 in biological fluids and can act as activity-based probes. Significance: These inhibitors will help clarify proteinase 3 function.

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Section: Discussionmentioning

confidence: 99%

New Selective Peptidyl Di(chlorophenyl) Phosphonate Esters for Visualizing and Blocking Neutrophil Proteinase 3 in Human Diseases

Guarino

Łęgowska

Epinette

et al. 2014

Journal of Biological Chemistry

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“…MCPR3-7 is a monoclonal mouse anti-PR3 antibody that can interfere with the catalytic activity of PR3 by an allosteric mechanism (Hinkofer et al, 2013). Initially, this mAb was generated to discriminate between proteolytically active mature PR3 and its inactive zymogen, which displays a different conformation.…”

Section: Protein and Chemical Inhibitors Of Proteinasementioning

confidence: 99%

“…MCPR3-7 also showed much stronger binding to the covalent PR3-a1PI complex than to the canonical PR3-a1PI complex. In the covalent PR3-a1PI complex, PR3 has a zymogen-like conformation that increases its affinity for MCPR3-7 (Hinkofer et al, 2013).…”

Section: Protein and Chemical Inhibitors Of Proteinasementioning

confidence: 99%

“…The binding of MCPR3-7 to PR3 also affects the covalent or canonical complexation with a1PI and delays the irreversible inhibition of PR3. MCPR3-7 is the first prototype of a PR3-directed conformation specific mAb with inhibitory properties (Hinkofer et al, 2013). Liu et al (2015) recently developed a novel strategy to design antibody-based inhibitors with nanomolar affinities for serine proteases with a trypsin-like fold (bovine trypsin and HNE) by engineering the exceptionally long CDR3H loop of a subgroup of bovine antibodies.…”

Section: Protein and Chemical Inhibitors Of Proteinasementioning

confidence: 99%

“…The inhibitory type of C-ANCA was most prevalent, but C-ANCAs with activity-enhancing effects on PR3 were also identified. C-ANCAs with inhibitory capacity partially block PR3 activity by an allosteric mechanism of inhibition (Hinkofer et al, 2013(Hinkofer et al, , 2015. Epitope mapping revealed that these mAbs bind mainly to a region that overlaps with that of group 1 (Hinkofer et al, unpublished data).…”

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Inhibitors and Antibody Fragments as Potential Anti-Inflammatory Therapeutics Targeting Neutrophil Proteinase 3 in Human Disease

Korkmaz¹,

Lesner²,

Guarino³

et al. 2016

Pharmacol Rev

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Proteinase 3 (PR3) has received great scientific attention after its identification as the essential antigenic target of antineutrophil cytoplasm antibodies in Wegener's granulomatosis (now called granulomatosis with polyangiitis). Despite many structural and functional similarities between neutrophil elastase (NE) and PR3 during biosynthesis, storage, and extracellular release, unique properties and pathobiological functions have emerged from detailed studies in recent years. The development of highly sensitive substrates and inhibitors of human PR3 and the creation of PR3-selective single knockout mice led to the identification of nonredundant roles of PR3 in cell death induction via procaspase-3 activation in cell cultures and in mouse models. According to a study in knockout mice, PR3 shortens the lifespan of infiltrating neutrophils in tissues and accelerates the clearance of aged neutrophils in mice. Membrane exposure of active human PR3 on apoptotic neutrophils reprograms the response of macrophages to phagocytosed neutrophils, triggers secretion of proinflammatory cytokines, and undermines immune silencing and tissue regeneration. PR3-induced disruption of the anti-inflammatory effect of efferocytosis may be relevant for not only granulomatosis with polyangiitis but also for other autoimmune diseases with high neutrophil turnover. Inhibition of membrane-bound PR3 by endogenous inhibitors such as the alpha-1-protease inhibitor is comparatively weaker than that of NE, suggesting that the adverse effects of unopposed PR3 activity resurface earlier than those of NE in individuals with alpha-1-protease inhibitor deficiency. Effective coverage of PR3 by anti-inflammatory tools and simultaneous inhibition of both PR3 and NE should be most promising in the future

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Thermophorese in Nanoliter‐Tropfen zur Quantifizierung von Aptamer‐Bindungen

et al. 2014

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Biomolekulare Interaktionen, die von zentraler Bedeutung für Pharmakologie und Diagnostik sind, können durch Thermophorese quantifiziert werden. Diese reagiert empfindlich auf bindungsinduzierte Größen‐, Ladungs‐ oder Konformationsänderungen. Etablierte Kapillarmessungen verbrauchen mindestens 0.5 μL pro Probe. Wir reduzierten den Probenverbrauch um den Faktor 50, indem wir 10‐nL‐Tropfen verwendeten, die mit akustischer Flüssigkeitsübertragung (Labcyte) hergestellt wurden. Die Tropfen wurden in einem Öl‐Tensid‐Gemisch stabilisiert und mit einem Infrarotlaser lokal erwärmt. Temperaturerhöhung, Marangoni‐Fluss und Konzentrationsverteilung wurden mit Fluoreszenzmikroskopie und numerischer Simulation analysiert. In 10‐nL‐Tropfen quantifizierten wir Affinität, Kooperativität und Pufferabhängigkeit des AMP‐Aptamers. Die Miniaturisierung und das 1536‐Well‐Platten‐Format machen die Methode hochdurchsatzfähig und automatisierbar. Dies forciert innovative Anwendungen in der serologischen Diagnostik und markierungsfreien Wirkstoffsuche.

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A Monoclonal Antibody (MCPR3-7) Interfering with the Activity of Proteinase 3 by an Allosteric Mechanism

Cited by 20 publications

References 37 publications

New Selective Peptidyl Di(chlorophenyl) Phosphonate Esters for Visualizing and Blocking Neutrophil Proteinase 3 in Human Diseases

New Selective Peptidyl Di(chlorophenyl) Phosphonate Esters for Visualizing and Blocking Neutrophil Proteinase 3 in Human Diseases

Inhibitors and Antibody Fragments as Potential Anti-Inflammatory Therapeutics Targeting Neutrophil Proteinase 3 in Human Disease

Thermophorese in Nanoliter‐Tropfen zur Quantifizierung von Aptamer‐Bindungen

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