A selective reversible azapeptide inhibitor of human neutrophil proteinase 3 derived from a high affinity FRET substrate

Autor: Martine Cadene, Christine Kellenberger, Brice Korkmaz, Christophe Epinette, Cécile Croix, Marie-Claude Viaud-Massuard, Lucie Jaquillard, Francis Gauthier, Gilles Lalmanach, Sylvain Marchand-Adam
Přispěvatelé: Pathologies Respiratoires : Protéolyse et Aérosolthérapie, Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique, immunothérapie, chimie et cancer (GICC), UMR 7292 CNRS [2012-2017] (GICC UMR 7292 CNRS), Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours-Centre National de la Recherche Scientifique (CNRS)
Rok vydání: 2011
Předmět:
Cathepsin G
Time Factors
MESH: Cathepsin G
MESH: Fluorescence Resonance Energy Transfer
Neutrophils
PATHOGENESIS
MESH: Flow Cytometry
MESH: Amino Acid Sequence
MESH: Neutrophils
MESH: Drug Design
Biochemistry
DISEASE
Substrate Specificity
chemistry.chemical_compound
0302 clinical medicine
MESH: Sputum
Proteinase 3
Fluorescence Resonance Energy Transfer
ACTIVE-SITE TITRANTS
MESH: Myeloblastin
Chromatography
High Pressure Liquid
0303 health sciences
biology
medicine.diagnostic_test
MESH: Kinetics
MESH: Peptides
Elastase
MESH: Proteinase Inhibitory Proteins
Secretory
Flow Cytometry
MESH: Leukocyte Elastase
030220 oncology & carcinogenesis
Neutrophil elastase
MESH: Oligopeptides
Oligopeptides
Protein Binding
Proteases
MESH: Pneumonia
Inhibitor
SURFACE
STRATEGIES
Proteolysis
Myeloblastin
Molecular Sequence Data
Proteinase Inhibitory Proteins
Secretory
Drug development
MESH: Proteolysis
Proteinase 3 (myeloblastin)
03 medical and health sciences
WEGENERS-GRANULOMATOSIS
medicine
MESH: Protein Binding
Humans
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
cardiovascular diseases
Amino Acid Sequence
MESH: Chromatography
High Pressure Liquid
Lung diseases
030304 developmental biology
Pharmacology
Serine protease
MESH: Humans
MESH: Molecular Sequence Data
MESH: Time Factors
Sputum
CATHEPSIN-G
Neutrophil extracellular traps
Pneumonia
Molecular biology
SERINE-PROTEASE
Kinetics
chemistry
Drug Design
Azapeptide
biology.protein
MESH: Substrate Specificity
HUMAN POLYMORPHONUCLEAR NEUTROPHILS
Leukocyte Elastase
Peptides
ELASTASE
Zdroj: Biochemical Pharmacology
Biochemical Pharmacology, Elsevier, 2012, 83 (6), pp.788-96. ⟨10.1016/j.bcp.2011.12.023⟩
ISSN: 1873-2968
0006-2952
DOI: 10.1016/j.bcp.2011.12.023⟩
Popis: International audience; The biological functions of human neutrophil proteinase 3 (PR3) remain unclear because of its close structural resemblance to neutrophil elastase and its apparent functional redundancy with the latter. Thus, all natural inhibitors of PR3 preferentially target neutrophil elastase. We have designed a selective PR3 inhibitor based on the sequence of one of its specific, sensitive FRET substrates. This azapeptide, azapro-3, inhibits free PR3 in solution, PR3 bound to neutrophil membranes, and the PR3 found in crude lung secretions from patients with chronic inflammatory pulmonary diseases. But it does not inhibit significantly neutrophil elastase or cathepsin G. Unlike most of azapeptides, this inhibitor does not form a stable acyl-enzyme complex; it is a reversible competitive inhibitor with a K(i) comparable to the K(m) of the parent substrate. Low concentrations (60 μM) of azapro-3 totally inhibited the PR3 secreted by triggered human neutrophils (200,000 cells/100 μL) and the PR3 in neutrophil homogenates and in lung secretions of patients with lung inflammation for hours. Azapro-3 also resisted proteolysis by all proteases contained in these samples for at least 2h.
Databáze: OpenAIRE
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