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-Kozicz, Jaak Jaeken, Paul Proost, Ghislain Opdenakker

Research output: Contribution to journalArticle

1 Citation (Scopus)

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-ATS 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)
JournalFEBS Journal
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Glycosylation
Galectin 3
Matrix Metalloproteinase 9
Matrix Metalloproteinase 3
Leukocyte Elastase
Chemical activation
Tiopronin
meprin A
Matrix Metalloproteinases
Oligosaccharides
Secreted Matrix Metalloproteinases
Matrix Metalloproteinase 12
Congenital Disorders of Glycosylation
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Proteolysis
Enzyme Precursors
Synovial Fluid
Physiology
Pathology
Gelatin

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Boon, L., Ugarte-Berzal, E., Martens, E., Vandooren, J., Rybakin, V., Colau, D., ... Opdenakker, G. (Accepted/In press). Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B. FEBS Journal. https://doi.org/10.1111/febs.14698

Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B. / Boon, Lise; Ugarte-Berzal, Estefania; Martens, Erik; Vandooren, Jennifer; Rybakin, Vasily; Colau, Didier; Gordon-Alonso, Monica; van der Bruggen, Pierre; Stöcker, Walter; Becker-Pauly, Christoph; Witters, Peter; Morava-Kozicz, Eva; Jaeken, Jaak; Proost, Paul; Opdenakker, Ghislain.

In: FEBS Journal, 01.01.2018.

Research output: Contribution to journalArticle

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-Kozicz, E, Jaeken, J, Proost, P & Opdenakker, G 2018, 'Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B', FEBS Journal. https://doi.org/10.1111/febs.14698
Boon, Lise ; Ugarte-Berzal, Estefania ; Martens, Erik ; Vandooren, Jennifer ; Rybakin, Vasily ; Colau, Didier ; Gordon-Alonso, Monica ; van der Bruggen, Pierre ; Stöcker, Walter ; Becker-Pauly, Christoph ; Witters, Peter ; Morava-Kozicz, Eva ; Jaeken, Jaak ; Proost, Paul ; Opdenakker, Ghislain. / Propeptide glycosylation and galectin-3 binding decrease proteolytic activation of human proMMP-9/progelatinase B. In: FEBS Journal. 2018.
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AU - Boon, Lise

AU - Ugarte-Berzal, Estefania

AU - Martens, Erik

AU - Vandooren, Jennifer

AU - Rybakin, Vasily

AU - Colau, Didier

AU - Gordon-Alonso, Monica

AU - van der Bruggen, Pierre

AU - Stöcker, Walter

AU - Becker-Pauly, Christoph

AU - Witters, Peter

AU - Morava-Kozicz, Eva

AU - Jaeken, Jaak

AU - Proost, Paul

AU - Opdenakker, Ghislain

PY - 2018/1/1

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N2 - 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-ATS 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.

AB - 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-ATS 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.

KW - galectin-3

KW - matrix metalloproteinase-9

KW - N-linked glycosylation

KW - propeptide

KW - proteolytic activation

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