A substrate-based approach to convert SerpinB1 into a specific inhibitor of proteinase 3, the Wegener's granulomatosis autoantigen

Autor: Christine Kellenberger, Hichem Lahouassa, Sandrine Castella, Elodie Pitois, Eileen Remold-O'Donnell, Brice Korkmaz, Gwenhael Jégot, Francis Gauthier, Chrystelle Derache, Marie Lise Jourdan
Přispěvatelé: 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)
Jazyk: angličtina
Rok vydání: 2011
Předmět:
Models
Molecular
Neutrophils
Protein Conformation
Cathepsin G
MESH: Neutrophils
Autoantigens
Biochemistry
MESH: Recombinant Proteins
chemistry.chemical_compound
MESH: Protein Conformation
Proteinase 3
SERINE PROTEINASES
ALPHA-1-PROTEASE INHIBITOR
MESH: Autoantibodies
CRYSTAL-STRUCTURE
Cloning
Molecular
Reactive center
IN-VIVO
MESH: Myeloblastin
0303 health sciences
biology
030302 biochemistry & molecular biology
Elastase
MESH: Wegener Granulomatosis
SERPINB1
Recombinant Proteins
3. Good health
Elastase inhibitor
MESH: Autoantigens
Neutrophil elastase
MESH: Models
Molecular
Biotechnology
MESH: Mutation
SURFACE
Myeloblastin
ALPHA-1-ANTITRYPSIN DEFICIENCY
03 medical and health sciences
MESH: Serpins
Genetics
Humans
MESH: Cloning
Molecular
cardiovascular diseases
Molecular Biology
Serpins
Autoantibodies
030304 developmental biology
Serine protease
CYSTIC-FIBROSIS
MESH: Humans
Granulomatosis with Polyangiitis
CATHEPSIN-G
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
chemistry
Mutation
biology.protein
ELASTASE INHIBITOR
HUMAN POLYMORPHONUCLEAR NEUTROPHILS
Zdroj: FASEB Journal
FASEB Journal, Federation of American Society of Experimental Biology, 2011, 25 (9), pp.3019-31. ⟨10.1096/fj.10-176552⟩
ISSN: 0892-6638
1530-6860
DOI: 10.1096/fj.10-176552⟩
Popis: International audience; The physiological and pathological functions of proteinase 3 (PR3) are not well understood due to its close similarity to human neutrophil elastase (HNE) and the lack of a specific inhibitor. Based on structural analysis of the active sites of PR3 and HNE, we generated mutants derived from the polyvalent inhibitor SerpinB1 (monocyte/neutrophil elastase inhibitor) that specifically inhibit PR3 and that differ from wt-SerpinB1 by only 3 or 4 residues in the reactive center loop. The rate constant of association between the best SerpinB1 mutant and PR3 is 1.4 × 10⁷ M⁻¹ * s⁻¹, which is ∼100-fold higher than that observed with wt-SerpinB1 and compares with that of α1-protease inhibitor (α1-PI) toward HNE. SerpinB1(S/DAR) is cleaved by HNE, but due to differences in rate, inhibition of PR3 by SerpinB1(S/DAR) is only minimally affected by the presence of HNE even when the latter is in excess. SerpinB1(S/DAR) inhibits soluble PR3 and also membrane-bound PR3 at the surface of activated neutrophils. Moreover, SerpinB1(S/DAR) clears induced PR3 from the surface of activated neutrophils. Overall, these specific inhibitors of PR3 will be valuable for defining biological functions of the protease and may prove useful as therapeutics for PR3-related inflammatory diseases, such as Wegener's granulomatosis.
Databáze: OpenAIRE
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