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

Scilabra, Simone D, Yamamoto, Kazuhiro, Pigoni, Martina, Sakamoto, Kazuma, Müller, Stephan A, Papadopoulou, Alkmini, Lichtenthaler, Stefan F, Troeberg, Linda ORCID: https://orcid.org/0000-0003-0939-4651, Nagase, Hideaki and Kadomatsu, Kenji (2017) 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. Matrix Biology, 59. pp. 69-79. ISSN 0945-053X

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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.

Item Type: Article
Additional Information: Copyright © 2016 Elsevier B.V. All rights reserved.
Uncontrolled Keywords: animals,binding sites,cos cells,cell line, tumor,cercopithecus aethiops,endocytosis,cytology,chemistry,gene expression regulation,hek293 cells,humans,kinetics,genetics,molecular sequence annotation,cytology,protein binding,protein interaction domains and motifs,protein interaction mapping,protein transport,genetics,genetics,signal transduction,solubility,genetics,transfection
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: LivePure Connector
Date Deposited: 09 Jan 2019 12:30
Last Modified: 21 Oct 2022 21:33
URI: https://ueaeprints.uea.ac.uk/id/eprint/69501
DOI: 10.1016/j.matbio.2016.07.004

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