%0 Journal Article
%A Scilabra, Simone D
%A Yamamoto, Kazuhiro
%A Pigoni, Martina
%A Sakamoto, Kazuma
%A Müller, Stephan A
%A Papadopoulou, Alkmini
%A Lichtenthaler, Stefan F
%A Troeberg, Linda
%A Nagase, Hideaki
%A Kadomatsu, Kenji
%T 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.
%J Matrix biology
%V 59
%@ 0945-053X
%C Amsterdam [u.a.]
%I Elsevier
%M DZNE-2020-05527
%P 69-79
%D 2017
%X 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.
%K Animals
%K Binding Sites
%K COS Cells
%K Cell Line, Tumor
%K Chlorocebus aethiops
%K Endocytosis
%K Epithelial Cells: cytology
%K Epithelial Cells: metabolism
%K Extracellular Matrix: chemistry
%K Extracellular Matrix: metabolism
%K Gene Expression Regulation
%K HEK293 Cells
%K Humans
%K Kinetics
%K Low Density Lipoprotein Receptor-Related Protein-1: genetics
%K Low Density Lipoprotein Receptor-Related Protein-1: metabolism
%K Molecular Sequence Annotation
%K Neuroglia: cytology
%K Neuroglia: metabolism
%K Protein Binding
%K Protein Interaction Domains and Motifs
%K Protein Interaction Mapping
%K Protein Transport
%K Receptors, Artificial: genetics
%K Receptors, Artificial: metabolism
%K Recombinant Proteins: genetics
%K Recombinant Proteins: metabolism
%K Signal Transduction
%K Solubility
%K Tissue Inhibitor of Metalloproteinase-3: genetics
%K Tissue Inhibitor of Metalloproteinase-3: metabolism
%K Transfection
%K LRP1 protein, human (NLM Chemicals)
%K Low Density Lipoprotein Receptor-Related Protein-1 (NLM Chemicals)
%K Receptors, Artificial (NLM Chemicals)
%K Recombinant Proteins (NLM Chemicals)
%K TIMP3 protein, human (NLM Chemicals)
%K Tissue Inhibitor of Metalloproteinase-3 (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:27476612
%R 10.1016/j.matbio.2016.07.004
%U https://pub.dzne.de/record/139205