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, 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
UEA Research Groups:	Faculty of Medicine and Health Sciences > Research Groups > Musculoskeletal Medicine Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health
Related URLs:	https://ora.ox.ac.uk/objects/uuid:ae875d...
Depositing User:	LivePure Connector
Date Deposited:	09 Jan 2019 12:30
Last Modified:	06 Feb 2025 08:11
URI:	https://ueaeprints.uea.ac.uk/id/eprint/69501
DOI:	10.1016/j.matbio.2016.07.004

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