1. New insights into the substrate specificity of macrophage elastase MMP-12
- Author
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Luiz Juliano, Anni Laffitte, Anne-Sophie Lamort, Rodolphe Gravier, Thierry Moreau, Marie-Louise Zani, Equipe Mecanismes Proteolyt Inflammat (CEPR INSERM U1100), Université de Tours, Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Universidade de São Paulo (USP), Conseil Régional Centre-Val de Loire Programme Investissement d'Avenir Grant Agreement Labex Mab'Improve ANR-10-LABX-53, ANR: ANR-10-LABX-53,programme Investissement d'Avenir Grant Agreement Labex Mab'Improve, Equipe Mecanismes Proteolyt Inflammat ( CEPR INSERM U1100 ), Université Tours, Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ), Universidade de São Paulo ( USP ), ANR : ANR-10-LABX-53,programme Investissement d'Avenir Grant Agreement Labex Mab'Improve, université de Bourgogne, CSGA, Laboratoires d'excellence - Optimization of therapeutic monoclonal antibodies development Better antibodies, better developed AND better used - - MAbImprove2010 - ANR-10-LABX-0053 - LABX - VALID, Centre d’Etude des Pathologies Respiratoires (CEPR), UMR 1100 (CEPR), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), Universidade de São Paulo = University of São Paulo (USP), ANR-10-LABX-0053,MAbImprove,Optimization of therapeutic monoclonal antibodies development Better antibodies, better developed AND better used(2010), Centre d'Etude des Pathologies Respiratoires (CEPR), UMR 1100. Equipe 2 'Mécanismes Protéolytiques dans l'Inflammation' (CEPR. Equipe 2), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), and Centre National de la Recherche Scientifique (CNRS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)
- Subjects
0301 basic medicine ,Clinical Biochemistry ,Organic chemistry ,Peptide ,Matrix metalloproteinase ,Bioinformatics ,Biochemistry ,enzyme specificity ,macrophage elastase ,mmp-12 ,peptide-protein docking ,substrate recognition ,Substrate Specificity ,Catalytic Domain ,Integral membrane protein ,chemistry.chemical_classification ,biology ,Biologie du développement ,Hemopexin ,Development Biology ,peptide ,integral membrane-proteins ,Molecular Docking Simulation ,emphysema ,MMP-12 ,Oligopeptides ,mice ,Structural similarity ,matrix metalloproteinase-12 ,Macrophage elastase ,03 medical and health sciences ,Species Specificity ,Matrix Metalloproteinase 12 ,expression ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Animals ,Humans ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,[ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Molecular Biology ,Fluorescent Dyes ,human alpha-1-proteinase inhibitor ,copd ,alveolar macrophages ,Chimie organique ,030104 developmental biology ,chemistry ,Docking (molecular) ,Biocatalysis ,biology.protein ,obstructive pulmonary-disease ,Elastin - Abstract
Macrophage elastase, or MMP-12, is mainly produced by alveolar macrophages and is believed to play a major role in the development of chronic obstructive pulmonary disease (COPD). The catalytic domain of MMP-12 is unique among MMPs in that it is very highly active on numerous substrates including elastin. However, measuring MMP-12 activity in biological fluids has been hampered by the lack of highly selective substrates. We therefore synthesized four series of fluorogenic peptide substrates based on the sequences of MMP-12 cleavage sites in its known substrates. Human MMP-12 efficiently cleaved peptide substrates containing a Pro at P3 in the sequence Pro-X-X↓Leu but lacked selectivity towards these substrates compared to other MMPs, including MMP-2, MMP-7, MMP-9 and MMP-13. On the contrary, the substrate Abz-RNALAVERTAS-EDDnp derived from the CXCR5 chemokine was the most selective substrate for MMP-12 ever reported. All substrates were cleaved more efficiently by full-length MMP-12 than by its catalytic domain alone, indicating that the C-terminal hemopexin domain influences substrate binding and/or catalysis. Docking experiments revealed unexpected interactions between the peptide substrate Abz-RNALAVERTAS-EDDn and MMP-12 residues. Most of our substrates were poorly cleaved by murine MMP-12 suggesting that human and murine MMP-12 have different substrate specificities despite their structural similarity.
- Published
- 2016
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