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Matrix Biol. 2017 May;59:69-79. doi: 10.1016/j.matbio.2016.07.004. Epub 2016 Jul 29.

Dissecting the interaction between tissue inhibitor of metalloproteinases-3 (TIMP-3) and low density lipoprotein receptor-related protein-1 (LRP-1): Development of a "TRAP" to increase levels of TIMP-3 in the tissue.

Author information

1
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan; Department of Neuroproteomics, Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Munich, Germany; Neuroproteomics, Klinikum rechts der Isar and Institute for Advanced Study, Technische Universität München, Munich, Germany. Electronic address: simone.scilabra@dzne.de.
2
Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford, United Kingdom.
3
Department of Neuroproteomics, Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Munich, Germany.
4
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
5
Department of Neuroproteomics, Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Munich, Germany; Neuroproteomics, Klinikum rechts der Isar and Institute for Advanced Study, Technische Universität München, Munich, Germany; Munich Cluster for Systems Neurology (SyNergy), Ludwig-Maximilians-University, Munich, Germany.

Abstract

Tissue inhibitor of metalloproteinases 3 (TIMP-3) is a key regulator of extracellular matrix turnover for its ability to inhibit matrix metalloproteinases (MMPs), adamalysin-like metalloproteinases (ADAMs) and ADAMs with thrombospondin motifs (ADAMTSs). TIMP-3 is a secreted protein whose extracellular levels are regulated by endocytosis via the low-density-lipoprotein receptor-related protein-1 (LRP-1). In this study we developed a molecule able to "trap" TIMP-3 extracellularly, thereby increasing its tissue bioavailability. LRP-1 contains four ligand-binding clusters. In order to investigate the TIMP-3 binding site on LRP-1, we generated soluble minireceptors (sLRPs) containing the four distinct binding clusters or part of each cluster. We used an array of biochemical methods to investigate the binding of TIMP-3 to different sLRPs. We found that TIMP-3 binds to the ligand-binding cluster II of the receptor with the highest affinity and a soluble minireceptor containing the N-terminal half of cluster II specifically blocked TIMP-3 internalization, without affecting the turnover of metalloproteinases. Mass spectrometry-based secretome analysis showed that this minireceptor, named T3TRAP, selectively increased TIMP-3 levels in the extracellular space and inhibited constitutive shedding of a number of cell surface proteins. In conclusion, T3TRAP represents a biological tool that can be used to modulate TIMP-3 levels in the tissue and could be potentially developed as a therapy for diseases characterized by a deficit of TIMP-3, including arthritis.

KEYWORDS:

ADAMTSs; ADAMs; Extracellular matrix; Inflammation; Low-density lipoprotein receptor-related protein-1; Matrix metalloproteinases; Tissue inhibitor of metalloproteinase-3

PMID:
27476612
DOI:
10.1016/j.matbio.2016.07.004
[Indexed for MEDLINE]

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