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Homotrimeric MMP-9 is an active hitchhiker on alpha-2-macroglobulin partially escaping protease inhibition and internalization through LRP-1

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Abstract

Proteolysis is a crucial process in life, tightly controlled by numerous natural protease inhibitors. In human blood, alpha-2-macroglobulin is an emergency protease inhibitor preventing coagulation and damage to endothelia and leukocytes. With the use of a unique protease trapping mechanism, alpha-2-macroglobulin lures active proteases into its snap-trap, shields these from potential substrates and ‘flags’ their complex for elimination by receptor-mediated endocytosis. Matrix metalloprotease-9/gelatinase B is a secreted protease increased in blood of patients with inflammations, vascular disorders and cancers. Matrix metalloprotease-9 occurs as monomers and stable homotrimers, but the reason for their co-existence remains obscure. We discovered that matrix metalloprotease-9 homotrimers undergo reduced anti-proteolytic regulation by alpha-2-macroglobulin and are able to travel as a proteolytically active hitchhiker on alpha-2-macroglobulin. As a comparison, we revealed that monomeric active matrix metalloprotease-9 is efficiently trapped by human plasma alpha-2-macroglobulin and this masks the detection of activated matrix metalloprotease-9 with standard analysis techniques. In addition, we show that alpha-2-macroglobulin/trimer complexes escape clearance through the receptor low-density lipoprotein receptor-related protein 1, also known as the alpha-2-macroglobulin receptor. Thus, the biochemistry and biology of matrix metalloprotease-9 monomers and trimers are completely different as multimerization enables active matrix metalloprotease-9 to partially avoid alpha-2-macroglobulin regulation both by direct protease inhibition and by removal from the extracellular space by receptor-mediated endocytosis. Finally, for the biomarker field, the analysis of alpha-2-macroglobulin/protease complexes with upgraded technology is advocated as a quotum for protease activation in human plasma samples.

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Abbreviations

α2M:

Alpha-2-macroglobulin

cdMMP-3:

Catalytic domain of MMP-3

LRP-1:

Low density lipoprotein receptor-related protein-1

MMPs:

Matrix metalloproteases

TIMPs:

Tissue inhibitors of metalloproteases

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Acknowledgements

The authors would like to thank Erik Martens, Karen Turneer, Jens Van Bael and Michael Van Canneyt for their assistance with the zymography analysis and the separation of MMP-9 monomers and trimers. EUB and JV are postdoctoral fellows funded by the Rega Foundation and the Research Foundation of Flanders (FWO-Vlaanderen). Project funding was from KU Leuven through a C1 grant (C16/17/010), the Belgian Charcot Foundation and FWO-Vlaanderen (Grants G0A7516N and G0A5716N).

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  1. Laboratory of Immunobiology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, KU Leuven, Herestraat 49, Bus 1044, 3000, Leuven, Belgium

    Xena Serifova, Estefania Ugarte-Berzal, Ghislain Opdenakker & Jennifer Vandooren

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  1. Xena Serifova
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  2. Estefania Ugarte-Berzal
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  3. Ghislain Opdenakker
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Serifova, X., Ugarte-Berzal, E., Opdenakker, G. et al. Homotrimeric MMP-9 is an active hitchhiker on alpha-2-macroglobulin partially escaping protease inhibition and internalization through LRP-1. Cell. Mol. Life Sci. 77, 3013–3026 (2020). https://doi.org/10.1007/s00018-019-03338-4

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