The cancer-associated meprin β variant G32R provides an additional activation site and promotes cancer cell invasion

Schäffler, Henning; Li, Wenjia; Helm, Ole; Krüger, Sandra; Böger, Christine; Peters, Florian; Röcken, Christoph; Sebens, Susanne; Lucius, Ralph; Becker‐Pauly, Christoph; Arnold, Philipp

doi:10.1242/jcs.220665

Cited by 12 publications

(9 citation statements)

References 33 publications

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“…Although the difference was not huge, an alternation of the proteolytic activity of meprin β likely affects the invasiveness of tumor cells. This result was in line with the results of the G32R variant (Schäffler et al, 2019), thus indicating a possible pro-metastatic function of meprin β.…”

Section: Discussionsupporting

confidence: 90%

“…This was, indeed, true for the basal activity; however, we did not observe increased proteolytic activity on cells expressing the G45R variant when incubated with trypsin. In studies investigating a meprin β G32R variant, the additional arginine, indeed, resulted in increased cell surface activity of meprin β (Schäffler et al, 2019). Analyzing the proteolytic processing of protein substrates, G45R showed a tendency of increased activity compared to wild-type meprin β.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the Cancer-Associated Meprin Βeta Variants G45R and G89R

Gellrich

Scharfenberg

Peters

et al. 2021

Front. Mol. Biosci.

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Meprin β is a metalloprotease associated with neurodegeneration, inflammation, extracellular matrix homeostasis, transendothelial cell migration, and cancer. In this study, we investigated two melanoma-associated variants of meprin β, both exhibiting a single amino acid exchange, namely, meprin β G45R and G89R. Based on the structural data of meprin β and with regard to the position of the amino acid exchanges, we hypothesized an increase in proteolytic activity in the case of the G45R variant due to the induction of a potential new activation site and a decrease in proteolytic activity from the G89R variant due to structural instability. Indeed, the G89R variant showed, overall, a reduced expression level compared to wild-type meprin β, accompanied by decreased activity and lower cell surface expression but strong accumulation in the endoplasmic reticulum. This was further supported by the analysis of the shedding of the interleukin-6 receptor (IL-6R) by meprin β and its variants. In transfected HEK cells, the G89R variant was found to generate less soluble IL-6R, whereas the expression of meprin β G45R resulted in increased shedding of the IL-6R compared to wild-type meprin β and the G89R variant. A similar tendency of the induced shedding capacity of G45R was seen for the well-described meprin β substrate CD99. Furthermore, employing an assay for cell migration in a collagen IV matrix, we observed that the transfection of wild-type meprin β and the G45R variant resulted in increased migration of HeLa cells, while the G89R variant led to diminished mobility.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Characterization of the Cancer-Associated Meprin Βeta Variants G45R and G89R

Gellrich

Scharfenberg

Peters

et al. 2021

Front. Mol. Biosci.

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“…Furthermore, the isoform meprin β mRNA, containing an additional 5 sequence, was discovered in human colon adenocarcinoma cell line (HT29-18C1) (Dietrich et al, 1996), human breast cancer cell lines (MCF-7 and SK-BR-3), human osteosarcoma cell line (U-2 OS), and the human pancreatic cancer cell line (BxPC-3), indicating its aspect in cancer (Matters and Bond, 1999). We could also show that single amino acid exchange variants of meprin β identified in cancer patients alter its proteolytic activity on the cell surface (Schäffler et al, 2019). In addition to direct cell-cell interaction, communication between cells can also occur through extracellular vesicles (EVs), which carry proteins like CD109 (Sakakura et al, 2016;Yamamoto et al, 2019).…”

Section: Introductionmentioning

confidence: 66%

Cell Surface Processing of CD109 by Meprin β Leads to the Release of Soluble Fragments and Reduced Expression on Extracellular Vesicles

Lückstädt¹,

Bub²,

Koudelka³

et al. 2021

Front. Cell Dev. Biol.

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Cluster of differentiation 109 (CD109) is a glycosylphosphatidylinositol (GPI)-anchored protein expressed on primitive hematopoietic stem cells, activated platelets, CD4+ and CD8+ T cells, and keratinocytes. In recent years, CD109 was also associated with different tumor entities and identified as a possible future diagnostic marker linked to reduced patient survival. Also, different cell signaling pathways were proposed as targets for CD109 interference including the TGFβ, JAK-STAT3, YAP/TAZ, and EGFR/AKT/mTOR pathways. Here, we identify the metalloproteinase meprin β to cleave CD109 at the cell surface and thereby induce the release of cleavage fragments of different size. Major cleavage was identified within the bait region of CD109 residing in the middle of the protein. To identify the structural localization of the bait region, homology modeling and single-particle analysis were applied, resulting in a molecular model of membrane-associated CD109, which allows for the localization of the newly identified cleavage sites for meprin β and the previously published cleavage sites for the metalloproteinase bone morphogenetic protein-1 (BMP-1). Full-length CD109 localized on extracellular vesicles (EVs) was also identified as a release mechanism, and we can show that proteolytic cleavage of CD109 at the cell surface reduces the amount of CD109 sorted to EVs. In summary, we identified meprin β as the first membrane-bound protease to cleave CD109 within the bait region, provide a first structural model for CD109, and show that cell surface proteolysis correlates negatively with CD109 released on EVs.

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“…Prior to functional tests, we further analyzed the stability of the full-length receptor on EVs, and therefore, incubated the purified vesicles for 24 h at 37 • C in PBS ( Figure 1E). No obvious cleavage fragments appeared over 24 h neither at~70 kDa (typical for ADAM proteases) [29,30] nor at~55 kDa, typical for meprin proteases and cancer-associated variants of meprin β ( Figure 1E) [22,31].…”

Section: Full-length Il-6 Receptor Is Present On Extracellular Vesiclmentioning

confidence: 99%

Joint Reconstituted Signaling of the IL-6 Receptor via Extracellular Vesicles

Arnold

Lückstädt

et al. 2020

Cells

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Interleukin-6 (IL-6) signaling is a crucial regulatory event important for many biological functions, such as inflammation and tissue regeneration. Accordingly, several pathological conditions are associated with dysregulated IL-6 activity, making it an attractive therapeutic target. For instance, blockade of IL-6 or its α-receptor (IL-6R) by monoclonal antibodies has been successfully used to treat rheumatoid arthritis. However, based on different signaling modes, IL-6 function varies between pro- and anti-inflammatory activity, which is critical for therapeutic intervention. So far, three modes of IL-6 signaling have been described, the classic anti-inflammatory signaling, as well as pro-inflammatory trans-signaling, and trans-presentation. The IL-6/IL-6R complex requires an additional β-receptor (gp130), which is expressed on almost all cells of the human body, to induce STAT3 (signal transducer and activator of signal transcription 3) phosphorylation and subsequent transcriptional regulation. In contrast, the IL-6R is expressed on a limited number of cells, including hepatocytes and immune cells. However, the proteolytic release of the IL-6R enables trans-signaling on cells expressing gp130 only. Here, we demonstrate a fourth possibility of IL-6 signaling that we termed joint reconstituted signaling (JRS). We show that IL-6R on extracellular vesicles (EVs) can also be transported to and fused with other cells that lack the IL-6R on their surface. Importantly, JRS via EVs induces delayed STAT3 phosphorylation compared to the well-established trans-signaling mode. EVs isolated from human serum were already shown to carry the IL-6R, and thus this new signaling mode should be considered with regard to signal intervention.

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The cancer-associated meprin β variant G32R provides an additional activation site and promotes cancer cell invasion

Cited by 12 publications

References 33 publications

Characterization of the Cancer-Associated Meprin Βeta Variants G45R and G89R

Characterization of the Cancer-Associated Meprin Βeta Variants G45R and G89R

Cell Surface Processing of CD109 by Meprin β Leads to the Release of Soluble Fragments and Reduced Expression on Extracellular Vesicles

Joint Reconstituted Signaling of the IL-6 Receptor via Extracellular Vesicles

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