TIMP-3 facilitates binding of target metalloproteinases to the endocytic receptor LRP-1 and promotes scavenging of MMP-1.

Published on Jul 21, 2020in Scientific Reports3.998
· DOI :10.1038/S41598-020-69008-9
Anna Paola Carreca7
Estimated H-index: 7
Veronica M. Pravata4
Estimated H-index: 4
(Dund.: University of Dundee)
+ 5 AuthorsSimone D. Scilabra8
Estimated H-index: 8
Matrix metalloproteinases (MMPs) and the related families of disintegrin metalloproteinases (ADAMs) and ADAMs with thrombospondin repeats (ADAMTSs) play a crucial role in extracellular matrix (ECM) turnover and shedding of cell-surface molecules. The proteolytic activity of metalloproteinases is post-translationally regulated by their endogenous inhibitors, known as tissue inhibitors of metalloproteinases (TIMPs). Several MMPs, ADAMTSs and TIMPs have been reported to be endocytosed by the low-density lipoprotein receptor-related protein-1 (LRP-1). Different binding affinities of these proteins for the endocytic receptor correlate with different turnover rates which, together with differences in their mRNA expression, determines their nett extracellular levels. In this study, we used surface plasmon resonance to evaluate the affinity between LRP-1 and a number of MMPs, ADAMs, ADAMTSs, TIMPs and metalloproteinase/TIMP complexes. This identified MMP-1 as a new LRP-1 ligand. Among the proteins analyzed, TIMP-3 bound to LRP-1 with highest affinity (KD = 1.68 nM). Additionally, we found that TIMP-3 can facilitate the clearance of its target metalloproteinases by bridging their binding to LRP-1. For example, the free form of MMP-1 was found to have a KD of 34.6 nM for LRP-1, while the MMP-1/TIMP-3 complex had a sevenfold higher affinity (KD = 4.96 nM) for the receptor. TIMP-3 similarly bridged binding of MMP-13 and MMP-14 to LRP-1. TIMP-1 and TIMP-2 were also found to increase the affinity of target metalloproteinases for LRP-1, albeit to a lesser extent. This suggests that LRP-1 scavenging of TIMP/metalloproteinase complexes may be a general mechanism by which inhibited metalloproteinases are removed from the extracellular environment.
📖 Papers frequently viewed together
14 Citations
78 Citations
#1Simone D. Scilabra (TUM: Technische Universität München)H-Index: 8
#2Martina Pigoni (German Center for Neurodegenerative Diseases)H-Index: 7
Last. Stefan F. LichtenthalerH-Index: 43
view all 7 authors...
The tissue inhibitor of metalloproteinases-3 (TIMP-3) is a major regulator of extracellular matrix turnover and protein shedding by inhibiting different classes of metalloproteinases, including disintegrin metalloproteinases (ADAMs). Tissue bioavailability of TIMP-3 is regulated by the endocytic receptor low-density-lipoprotein receptor-related protein-1 (LRP-1). TIMP-3 plays protective roles in disease. Thus, different approaches have been developed aiming to increase TIMP-3 bioavailability, ye...
13 CitationsSource
#1Simone D. Scilabra (Nagoya University)H-Index: 8
#2Kazuhiro Yamamoto (University of Oxford)H-Index: 17
Last. Kenji Kadomatsu (Nagoya University)H-Index: 74
view all 10 authors...
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 increasi...
14 CitationsSource
#1Kazuhiro Yamamoto (University of Oxford)H-Index: 17
#2Hiroshi Okano (University of Oxford)H-Index: 1
Last. Hideaki Nagase (University of Oxford)H-Index: 103
view all 11 authors...
Matrix metalloproteinase 13 (MMP-13) degrades collagenous extracellular matrix and its aberrant activity associates with diseases such as arthritis, cancer, atherosclerosis and fibrosis. The wide range of MMP-13 proteolytic capacity suggests that it is a powerful, potentially destructive proteinase and thus it has been believed that MMP-13 is not produced in most adult human tissues in the steady state. Present study has revealed that human chondrocytes isolated from healthy adults constitutivel...
80 CitationsSource
#1Christine Doherty (Arthritis Research UK)H-Index: 3
#2Robert Visse (Arthritis Research UK)H-Index: 19
Last. Linda Troeberg (Arthritis Research UK)H-Index: 28
view all 6 authors...
Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a central inhibitor of matrix-degrading and sheddase families of metalloproteinases. Extracellular levels of the inhibitor are regulated by the balance between its retention on the extracellular matrix and its endocytic clearance by the scavenger receptor low density lipoprotein receptor-related protein 1 (LRP1). Here, we used molecular modeling to predict TIMP-3 residues potentially involved in binding to LRP1 based on the proposed LRP1 bindi...
14 CitationsSource
#1Jessica Thevenard (CNRS: Centre national de la recherche scientifique)H-Index: 9
#2L. Verzeaux (CNRS: Centre national de la recherche scientifique)H-Index: 4
Last. Hervé Emonard (CNRS: Centre national de la recherche scientifique)H-Index: 33
view all 15 authors...
Tissue inhibitor of metalloproteinases-1 (TIMP-1) regulates the extracellular matrix turnover by inhibiting the proteolytic activity of matrix metalloproteinases (MMPs). TIMP-1 also displays MMP-independent activities that influence the behavior of various cell types including neuronal plasticity, but the underlying molecular mechanisms remain mostly unknown. The trans-membrane receptor low-density lipoprotein receptor-related protein-1 (LRP-1) consists of a large extracellular chain with distin...
25 CitationsSource
#1David Marchant (UBC: University of British Columbia)H-Index: 12
#2Caroline L. Bellac (UBC: University of British Columbia)H-Index: 10
Last. Christopher M. OverallH-Index: 99
view all 26 authors...
Matrix metalloproteinases (MMPs) normally act extracellularly. Now Marchant et al. report an unexpected nuclear activity for MMP-12 in virus-infected cells in regulating transcription of the gene encoding IκBα and affecting secretion of interferon-α.
178 CitationsSource
#1Kazuhiro Yamamoto (University of Oxford)H-Index: 17
#2Kathryn Owen (Imperial College London)H-Index: 1
Last. Hideaki Nagase (University of Oxford)H-Index: 103
view all 8 authors...
Degradation of the cartilage proteoglycan aggrecan is an early event in the development of osteoarthritis, and a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) and ADAMTS-5 are considered to be the major aggrecan-degrading enzymes. We have recently found that ADAMTS-5 is rapidly endocytosed via low density lipoprotein receptor-related protein 1 (LRP1) and degraded by chondrocytes. Here we report that this regulatory mechanism also applies to ADAMTS-4, although its rate...
54 CitationsSource
#1Enrico A. Stura (Commissariat à l'énergie atomique et aux énergies alternatives)H-Index: 57
#2Robert Visse (University of Oxford)H-Index: 19
Last. Hideaki Nagase (University of Oxford)H-Index: 103
view all 5 authors...
Matrix metalloproteinase (MMP)-13 is one of the mammalian collagenases that play key roles in tissue remodelling and repair and in progression of diseases such as cancer, arthritis, atherosclerosis, and aneurysm. For collagenase to cleave triple helical collagens, the triple helical structure has to be locally unwound before hydrolysis, but this process is not well understood. We report crystal structures of catalytically inactive full-length human MMP-13(E223A) in complex with peptides of 14–26...
28 CitationsSource
#1Kazuhiro YamamotoH-Index: 17
#2Linda TroebergH-Index: 28
Last. Hideaki NagaseH-Index: 103
view all 7 authors...
Aggrecan is a major matrix component of articular cartilage, and its degradation is a crucial event in the development of osteoarthritis (OA). Adamalysin-like metalloproteinase with thrombospondin motifs 5 (ADAMTS-5) is a major aggrecan-degrading enzyme in cartilage, but there is no clear correlation between ADAMTS-5 mRNA levels and OA progression. Here, we report that post-translational endocytosis of ADAMTS-5 by chondrocytes regulates its extracellular activity. We found 2- to 3-fold reduced a...
67 CitationsSource
#1Simone D. Scilabra (University of Oxford)H-Index: 8
#2Linda Troeberg (University of Oxford)H-Index: 28
Last. Hideaki Nagase (University of Oxford)H-Index: 103
view all 8 authors...
Tissue inhibitor of metalloproteinases-3 (TIMP-3) plays a key role in regulating extracellular matrix turnover by inhibiting matrix metalloproteinases (MMPs), adamalysins (ADAMs), and adamalysins with thrombospondin motifs (ADAMTSs). We demonstrate that levels of this physiologically important inhibitor can be regulated post-translationally by endocytosis. TIMP-3 was endocytosed and degraded by a number of cell types including chondrocytes, fibroblasts, and monocytes, and we found that the endoc...
48 CitationsSource
Cited By5
#1Keron W. J. Rose (ISMMS: Icahn School of Medicine at Mount Sinai)
#2Nandaraj Taye (ISMMS: Icahn School of Medicine at Mount Sinai)H-Index: 11
Last. Dirk Hubmacher (ISMMS: Icahn School of Medicine at Mount Sinai)H-Index: 19
view all 4 authors...
A disintegrin and metalloprotease with thrombospondin type I motifs (ADAMTS) proteases are secreted metalloproteinases that play key roles in the formation, homeostasis and remodeling of the extracellular matrix (ECM). The substrate spectrum of ADAMTS proteases can range from individual ECM proteins to entire families of ECM proteins, such as the hyalectans. ADAMTS-mediated substrate cleavage is required for the formation, remodeling and physiological adaptation of the ECM to the needs of indivi...
1 CitationsSource
#1Anna Paola Carreca (ISMETT)H-Index: 7
#2Veronica M. Pravata (Dund.: University of Dundee)H-Index: 4
Last. Simone D. Scilabra (ISMETT)H-Index: 8
view all 9 authors...
Ectodomain shedding is a key mechanism of several biological processes, including cell-communication. Disintegrin and metalloproteinases (ADAMs), together with the membrane-type matrix metalloproteinases, play a pivotal role in shedding transmembrane proteins. Aberrant shedding is associated to several pathological conditions, including arthritis. Tissue inhibitor of metalloproteases 3 (TIMP-3), an endogenous inhibitor of ADAMs and matrix metalloproteases (MMPs), has been proven to be beneficial...
1 CitationsSource
#2Doretta Cuffaro (UniPi: University of Pisa)H-Index: 5
Last. Simone D. Scilabra (ISMETT)H-Index: 8
view all 7 authors...
For decades, disintegrin and metalloproteinase 17 (ADAM17) has been the object of deep investigation. Since its discovery as the tumor necrosis factor convertase, it has been considered a major drug target, especially in the context of inflammatory diseases and cancer. Nevertheless, the development of drugs targeting ADAM17 has been harder than expected. This has generally been due to its multifunctionality, with over 80 different transmembrane proteins other than tumor necrosis factor α (TNF) b...
7 CitationsSource
#1Oliver McClurg (UEA: University of East Anglia)
#2Ryan Tinson (UEA: University of East Anglia)H-Index: 4
Last. Linda Troeberg (UEA: University of East Anglia)H-Index: 28
view all 3 authors...
Osteoarthritis is a common, degenerative joint disease with significant socio-economic impact worldwide. There are currently no disease-modifying drugs available to treat the disease, making this an important area of pharmaceutical research. In this review, we assessed approaches being explored to directly inhibit metalloproteinase-mediated cartilage degradation and to counteract cartilage damage by promoting growth factor-driven repair. Metalloproteinase-blocking antibodies are discussed, along...
2 CitationsSource
#1Allison L. AraiH-Index: 3
#2Mary MiglioriniH-Index: 15
Last. Dudley K. StricklandH-Index: 80
view all 8 authors...
Matrix metalloprotease (MMP) activation contributes to degradation of the extracellular matrix (ECM), resulting in a multitude of pathologies. Low-density lipoprotein receptor-related protein 1 (LRP1) is a multifaceted endocytic and signaling receptor responsible for internalization and lysosomal degradation of diverse proteases, protease inhibitors and lipoproteins along with numerous other proteins. In the current study, we identified MMP-1 as a novel LRP1 ligand. Binding studies employing sur...
2 CitationsSource