Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B

Lise Boon, Estefania Ugarte-Berzal, Erik Martens, Jennifer Vandooren, Vasily Rybakin, Didier Colau, Monica Gordon-Alonso, Pierre van der Bruggen, Walter Stöcker, Christoph Becker-Pauly, Peter Witters, Eva Morava, Jaak Jaeken, Paul Proost, Ghislain Opdenakker

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Matrix metalloproteinases (MMPs) are secreted as proenzymes, containing propeptides that interact with the catalytic zinc, thereby controlling MMP activation. The MMP-9 propeptide is unique in the MMP family because of its post-translational modification with an N-linked oligosaccharide. ProMMP-9 activation by MMP-3 occurs stepwise by cleavage of the propeptide in an aminoterminal (pro-AT) and carboxyterminal (pro-CT) peptide. We chemically synthesized aglycosyl pro-AT and pro-CT and purified recombinant glycosylated pro-AT S f−9 . First, we report new cleavage sites in the MMP-9 propeptide by MMP-3 and neutrophil elastase. Additionally, we demonstrated with the use of western blot analysis a higher resistance of glycosylated versus aglycosyl pro-AT against proteolysis by MMP-3, MMP-9, meprin α, neutrophil elastase and by protease-rich synovial fluids from rheumatoid arthritis patients. Moreover, we investigated the effect of glycosylation on proteolytic activation of human proMMP-9 with the use of zymography and dye-quenched gelatin cleavage analysis. Compared to recombinant Sf-9 proMMP-9 glycoforms, larger oligosaccharides of human neutrophil proMMP-9 increased resistance against proteolytic activation. Additionally, proMMP-9 from Congenital Disorder of Glycosylation patients, compared to healthy controls, showed a higher activation rate by MMP-3. Finally, we demonstrated that glycan-galectin-3 interactions reduced proMMP-9 activation. In conclusion, modification of MMP-9 propeptide glycosylation is a fine-tuning mechanism and co-determines the specific activity of MMP-9 in physiology and pathology. Enzymes: MMP-9 EC 3.4.24.35, MMP-3 EC 3.4.24.17, meprin α EC 3.4.24.18, neutrophil elastase EC 3.4.21.37, trypsin EC 3.4.21.4 and PNGase F EC 3.5.1.52.

Original languageEnglish (US)
Pages (from-to)930-945
Number of pages16
JournalFEBS Journal
Volume286
Issue number5
DOIs
StatePublished - Mar 2019

Keywords

  • N-linked glycosylation
  • galectin-3
  • matrix metalloproteinase-9
  • propeptide
  • proteolytic activation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Boon, L., Ugarte-Berzal, E., Martens, E., Vandooren, J., Rybakin, V., Colau, D., Gordon-Alonso, M., van der Bruggen, P., Stöcker, W., Becker-Pauly, C., Witters, P., Morava, E., Jaeken, J., Proost, P., & Opdenakker, G. (2019). Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B. FEBS Journal, 286(5), 930-945. https://doi.org/10.1111/febs.14698