TY  - JOUR
AU  - Scilabra, Simone D
AU  - Yamamoto, Kazuhiro
AU  - Pigoni, Martina
AU  - Sakamoto, Kazuma
AU  - Müller, Stephan A
AU  - Papadopoulou, Alkmini
AU  - Lichtenthaler, Stefan F
AU  - Troeberg, Linda
AU  - Nagase, Hideaki
AU  - Kadomatsu, Kenji
TI  - 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.
JO  - Matrix biology
VL  - 59
SN  - 0945-053X
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - DZNE-2020-05527
SP  - 69-79
PY  - 2017
AB  - 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.
KW  - Animals
KW  - Binding Sites
KW  - COS Cells
KW  - Cell Line, Tumor
KW  - Chlorocebus aethiops
KW  - Endocytosis
KW  - Epithelial Cells: cytology
KW  - Epithelial Cells: metabolism
KW  - Extracellular Matrix: chemistry
KW  - Extracellular Matrix: metabolism
KW  - Gene Expression Regulation
KW  - HEK293 Cells
KW  - Humans
KW  - Kinetics
KW  - Low Density Lipoprotein Receptor-Related Protein-1: genetics
KW  - Low Density Lipoprotein Receptor-Related Protein-1: metabolism
KW  - Molecular Sequence Annotation
KW  - Neuroglia: cytology
KW  - Neuroglia: metabolism
KW  - Protein Binding
KW  - Protein Interaction Domains and Motifs
KW  - Protein Interaction Mapping
KW  - Protein Transport
KW  - Receptors, Artificial: genetics
KW  - Receptors, Artificial: metabolism
KW  - Recombinant Proteins: genetics
KW  - Recombinant Proteins: metabolism
KW  - Signal Transduction
KW  - Solubility
KW  - Tissue Inhibitor of Metalloproteinase-3: genetics
KW  - Tissue Inhibitor of Metalloproteinase-3: metabolism
KW  - Transfection
KW  - LRP1 protein, human (NLM Chemicals)
KW  - Low Density Lipoprotein Receptor-Related Protein-1 (NLM Chemicals)
KW  - Receptors, Artificial (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
KW  - TIMP3 protein, human (NLM Chemicals)
KW  - Tissue Inhibitor of Metalloproteinase-3 (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:27476612
DO  - DOI:10.1016/j.matbio.2016.07.004
UR  - https://pub.dzne.de/record/139205
ER  -