Biology subjects

Troeberg, L.

Publications and source records attributed to Troeberg, L..

4 recordsLinked to original sources

bioRxiv2025

Molecular determinants of extracellular TIMP-3 accumulation in Sorsby fundus dystrophy.

Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a critical regulator of extracellular matrix turnover. Mutations in TIMP-3 cause Sorsby fundus dystrophy (SFD), an inherited macular dystrophy that is similar to age-related macular degeneration (AMD) but which generally presents earlier. SFD is characterised by the accumulation of mutant TIMP-3 protein in Bruchs membrane, a multilaminar extracellular matrix underlying the retinal pigment epithelium (RPE). Here, we show that RPE cells regulate wild-type TIMP-3 levels post-translationally, with ARPE-19 and hTERT RPE-1 cell lines endocytosing the protein via the low-density lipoprotein receptor-related protein (LRP) family of scavenger receptors. LRP-mediated endocytosis of the SFD TIMP-3 variants S204C and Y191C was significantly delayed, establishing a molecular mechanism for their extracellular accumulation in SFD. In contrast, endocytosis of the SFD variant H181R TIMP-3 was unaltered, suggesting it accumulates through a distinct molecular mechanism, potentially via increased retention on extracellular matrix heparan sulfate proteoglycans. These findings reveal heterogeneity in the molecular mechanism of SFD pathogenesis, which has direct implications for therapeutic development. Genotype-specific interventions may be required, such as strategies that enhance receptor-mediated clearance or disrupt extracellular TIMP-3 retention. Our study also has broader implications for AMD, where TIMP-3 and other LRP and heparan sulfate ligands accumulate in drusen within Bruchs membrane. Age- and inflammation-dependent alterations in LRP expression and heparan sulfate structure may contribute to drusen formation and AMD progression. Understanding TIMP-3 trafficking in both physiological and pathological contexts could inform targeted treatments for SFD, AMD, and other degenerative disorders involving extracellular matrix dysregulation.

biochemistry↗

bioRxiv2024

The extracellular heparan sulfatase SULF2 limits myeloid IFNbeta signaling and Th17 responses in inflammatory arthritis.

Heparan sulfate (HS) proteoglycans are important regulators of cellular responses to soluble mediators such as chemokines, cytokines and growth factors. We profiled changes in expression of genes encoding HS core proteins, biosynthesis enzymes and modifiers during macrophage polarisation, and found that the most highly regulated gene was Sulf2, an extracellular HS 6-O-sulfatase that was markedly downregulated in response to pro-inflammatory stimuli. We then generated Sulf2+/- bone marrow chimeric mice and examined inflammatory responses in antigen-induced arthritis, as a model of rheumatoid arthritis. Resolution of inflammation was impaired in myeloid Sulf2+/- chimeras, with elevated joint swelling and increased abundance of pro-arthritic Th17 cells in synovial tissue. Transcriptomic and in vitro analyses indicated that Sulf2 deficiency increased type I interferon signaling in bone marrow-derived macrophages, leading to elevated expression of the Th17-inducing cytokine IL-6. This establishes that dynamic remodeling of HS by Sulf2 limits type I interferon signaling in macrophages, and so protects against Th17-driven pathology.

immunology↗

bioRxiv2021

Identification that ADAM17 mediates proteolytic maturation of calcium channel auxiliary α2δ subunits, and enables calcium current enhancement

The auxiliary 2{delta} subunits of voltage-gated calcium (CaV) channels are key to augmenting expression and function of CaV1 and CaV2 channels, and are also important drug targets in several therapeutic areas, including neuropathic pain. The 2{delta} proteins are translated as pre-proteins encoding both 2 and {delta}, and post-translationally proteolysed into 2 and {delta} subunits, which remain associated as a complex. In this study we have identified ADAM17 as a key protease involved in proteolytic processing of pro-2{delta}-1 and 2{delta}-3 subunits. We provide three lines of evidence: firstly, proteolytic cleavage is inhibited by chemical inhibitors of particular metalloproteases, including ADAM17. Secondly, proteolytic cleavage of both 2{delta}-1 and 2{delta}-3 is markedly reduced in cell lines by knockout of ADAM17 but not ADAM10. Thirdly, proteolytic cleavage is reduced by the N-terminal active domain of TIMP-3 (N-TIMP-3), which selectively inhibits ADAM17. We have found previously that proteolytic cleavage into mature 2{delta} is essential for the enhancement of CaV function, and in agreement, knockout of ADAM17 inhibited the ability of 2{delta}-1 to enhance both CaV2.2 and CaV1.2 calcium currents. Thus, our study identifies ADAM17 as a key protease required for proteolytic maturation of 2{delta}-1 and 2{delta}-3, and thus a potential drug target in neuropathic pain.

neuroscience↗

bioRxiv2019

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

Matrix metalloproteinases (MMPs) and the related families of disintegrin metalloproteinases (ADAMs) and ADAMs with thrombospondin repeats (ADAMTSs) play a crucial role in the turnover of extracellular matrix (ECM) components 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 analysed, 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 7-fold 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. These findings suggest that LRP-1 scavenging of TIMP/metalloproteinase complexes may be a general mechanism by which inhibited metalloproteinases are removed from the extracellular environment.

biochemistry↗