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New insights into the substrate specificity of macrophage elastase MMP-12

  • Anne-Sophie Lamort , Rodolphe Gravier , Anni Laffitte , Luiz Juliano , Marie-Louise Zani und Thierry Moreau EMAIL logo
Veröffentlicht/Copyright: 12. Januar 2016
Biological Chemistry
Aus der Zeitschrift Biological Chemistry Band 397 Heft 5

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.

Keywords: enzyme specificity; macrophage elastase; MMP-12; peptide-protein docking; substrate recognition

Acknowledgments:

Mass spectrometry analyses were performed by the ‘Plateforme de Spectrométrie de Masse et Protéomique’ of the Center for Molecular Biophysics in Orléans, France. N-terminal amino acid sequence analyses were performed by the ‘Plateforme Protéomique Pissaro’ at the University of Rouen, France. ASL was supported by a PhD fellowship from the Conseil Régional Centre-Val de Loire. RG was awarded with a post-doctoral fellowship from the Conseil Régional Centre-Val de Loire and the programme Investissement d’Avenir Grant Agreement Labex Mab’Improve (ANR-10-LABX-53). The English text was edited by Dr. Owen Parkes.

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Supplemental Material:

The online version of this article (DOI: 10.1515/hsz-2015-0254) offers supplementary material, available to authorized users.

Received: 2015-9-24
Accepted: 2016-1-4
Published Online: 2016-1-12
Published in Print: 2016-5-1

©2016 by De Gruyter

Artikel in diesem Heft

  1. Frontmatter
  2. Review
  3. Nrf2 activation in the treatment of neurodegenerative diseases: a focus on its role in mitochondrial bioenergetics and function
  4. Research Articles/Short Communications
  5. Protein Structure and Function
  6. Effect of molecular chaperones on aberrant protein oligomers in vitro: super-versus sub-stoichiometric chaperone concentrations
  7. Two new isoforms of the human hepatoma-derived growth factor interact with components of the cytoskeleton
  8. Cell Biology and Signaling
  9. Activation of corticotropin releasing factor receptors up regulates collagen production by hepatic stellate cells via promoting p300 expression
  10. On the regulative role of the glutamate receptor in mitochondria
  11. Proteolysis
  12. Biomechanical and biochemical regulation of cathepsin K expression in endothelial cells converge at AP-1 and NF-κB
  13. New insights into the substrate specificity of macrophage elastase MMP-12
https://doi.org/10.1515/hsz-2015-0254
Schlagwörter für diesen Artikel
enzyme specificity; macrophage elastase; MMP-12; peptide-protein docking; substrate recognition

Artikel in diesem Heft

  1. Frontmatter
  2. Review
  3. Nrf2 activation in the treatment of neurodegenerative diseases: a focus on its role in mitochondrial bioenergetics and function
  4. Research Articles/Short Communications
  5. Protein Structure and Function
  6. Effect of molecular chaperones on aberrant protein oligomers in vitro: super-versus sub-stoichiometric chaperone concentrations
  7. Two new isoforms of the human hepatoma-derived growth factor interact with components of the cytoskeleton
  8. Cell Biology and Signaling
  9. Activation of corticotropin releasing factor receptors up regulates collagen production by hepatic stellate cells via promoting p300 expression
  10. On the regulative role of the glutamate receptor in mitochondria
  11. Proteolysis
  12. Biomechanical and biochemical regulation of cathepsin K expression in endothelial cells converge at AP-1 and NF-κB
  13. New insights into the substrate specificity of macrophage elastase MMP-12
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