Cellular glycosylphosphatidylinositol-specific phospholipase D regulates urokinase receptor shedding and cell surface expression

Wilhelm, Olaf G.; Wilhelm, Sabine; Escott, Gemma M.; Lutz, Veronika; Magdolen, Viktor; Schmitt, Manfred; Rifkin, Daniel B.; Wilson, EL; Graeff, H.; Brunner, Georg

doi:10.1002/(sici)1097-4652(199908)180:2<225::aid-jcp10>3.0.co;2-2

Cited by 127 publications

(23 citation statements)

References 57 publications

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“…The source of such soluble CD87 species in human body fluids has therefore long been discussed. Whereas they can derive from tumor cells and non‐malignant stromal cells, as well as from inflammatory cells [1,5,17], the mechanisms for release can include exposure to pathogens or their products, such as PI‐PLC [17,36], and exposure to host enzymes such as PI‐PLD, known to be overexpressed in some cancers [28], and proteinases. Indeed, the susceptibility of the D1–D2 linker sequence of CD87 to various Ser‐proteinases, including uPA, plasmin, several MMP and leukocyte Ser‐proteinases [17,20,22], elucidates the mechanism for the release of free D1.…”

Section: Discussionmentioning

confidence: 99%

“…Accordingly, a soluble D1 domain can be detected in body fluids such as urine, and is markedly increased in pathological states associated with various types of carcinomas or infectious and inflammatory disorders [15,23,24]. However, soluble forms of full‐length (D1D2D3) or truncated (D2D3) CD87 species are observed in urine and in plasma, and are similarly increased in diseases [5,23], for which they are now proposed as prognostic markers [23–26], although mechanistic explanations for their release remain scarce [22,27,28].…”

Section: Introductionmentioning

confidence: 99%

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Plasmin cleaves the juxtamembrane domain and releases truncated species of the urokinase receptor (CD87) from human bronchial epithelial cells

Beaufort¹,

Leduc²,

Rousselle³

et al. 2004

FEBS Letters

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The three-domain (D1D2D3) urokinase receptor (CD87) is highly susceptible to cleavage within the D1-D2 linker sequence, but also within the juxtamembrane region by yet poorly characterized proteinases, allowing the release of D1 and D2D3 species in various (patho)physiological body fluids. Using immunoblot analysis and ELISA applied to a recombinant soluble CD87 and to CD87-expressing epithelial cells, we establish that exogenous or in situ generated plasmin proteolyzes CD87 in the D1-D2 linker and D3 carboxyterminal sequences, producing a major soluble D2D3 species. Mass spectrometry analysis of the fragmentation of CD87-related synthetic peptides, and aminoterminal sequencing of D2D3 reveal Arg 83 , Arg 89 , and Arg 281 as residues targeted by plasmin within human CD87.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Plasmin cleaves the juxtamembrane domain and releases truncated species of the urokinase receptor (CD87) from human bronchial epithelial cells

Beaufort¹,

Leduc²,

Rousselle³

et al. 2004

FEBS Letters

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“…The Ly‐6 proteins are normally glycosyl phosphatidylinositol (GPI)‐anchored cell surface, cysteine‐rich molecules, such as CD59 (Davies et al 1989), sperm acrosomal protein (SP‐10) (Palfree 1996), and the urokinase plasminogen activator (uPA) receptor (uPAR) (Wilhelm et al 1999; Stroncek et al 2004 and references therein). In addition, secreted members of the superfamily lacking the GPI anchor, such as the snake neurotoxins (Fleming et al 1993) and human SLURP‐1 (secreted Ly‐6/uPAR related protein 1) (Adermann et al 1999) and SLURP‐2 (Tsuji et al 2003), as well as secreted rat urinary proteins (Southan et al 2002) have also been described.…”

mentioning

confidence: 99%

Characterization of the five novel Ly‐6 superfamily members encoded in the MHC, and detection of cells expressing their potential ligands

Mallya¹,

Campbell²,

Aguado

2006

Protein Science

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Lymphocyte Antigen 6 (Ly-6) superfamily members are cysteine-rich, generally GPI-anchored cell surface proteins, which have definite or putative immune related roles. There are 27 members of this family described so far in the human genome and 37 in the mouse. Five of them are clustered in the class III region of the human and mouse MHCs. Following computational analyses, we functionally characterized the encoded proteins by creating epitope-tagged fusion constructs to determine molecular weight, complex formation, subcellular localization, post-translational modifications and ligand binding. We found that all human and mouse proteins were glycosylated, and most could form part of larger complexes. Human and mouse Ly6G6c and Ly6G6d, and mouse Ly6g6e were found to be GPIanchored cell surface proteins, highly expressed at the leading edges of cells, on filopodia, which are normally involved in cell adhesion and migration. However, analysis of Ly6G5c and Ly6G5b indicated that they are potentially secreted proteins. Our results indicate that there are two subclusters of related Ly-6 proteins in this region of the MHC, with Ly6G6c, Ly6G6d, and Ly6G6e forming one and Ly6G5c and Ly6G5b forming another. In addition, by FACS analysis we have found that the potential ligands for human LY6G6C, LY6G6D, and LY6G5C are expressed on K562 cells, an undifferentiated megakaryocyte cell line, indicating a potential role in hematopoietic cell differentiation. This characterization of the five MHC class III region Ly-6 family members is of great relevance, as they represent 18% of the human Ly-6 protein family and 50% of the secreted ones.

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“…SuPAR could not be detected in the concentrated conditioned medium from AT84-EV cells, nor in AT84-uPAR concentrated conditioned FBSM (Fig. 4b), as expected due to serum-induced inhibition of phospholipases [62, 63]. To test whether mouse suPAR could function as a chemoattractant as previously reported for human uPAR [64], the conditioned SFM from AT84-EV and AT84-uPAR cells was harvested and used as an attractant in a migration assay using the AT84-EV cells (Fig.…”

Section: Resultsmentioning

confidence: 55%

Cleavage of the urokinase receptor (uPAR) on oral cancer cells: regulation by transforming growth factor – β1 (TGF-β1) and potential effects on migration and invasion

et al. 2017

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BackgroundUrokinase plasminogen activator (uPA) receptor (uPAR) is up-regulated at the invasive tumour front of human oral squamous cell carcinoma (OSCC), indicating a role for uPAR in tumour progression. We previously observed elevated expression of uPAR at the tumour-stroma interface in a mouse model for OSCC, which was associated with increased proteolytic activity. The tumour microenvironment regulated uPAR expression, as well as its glycosylation and cleavage. Both full-length- and cleaved uPAR (uPAR (II-III)) are involved in highly regulated processes such as cell signalling, proliferation, migration, stem cell mobilization and invasion. The aim of the current study was to analyse tumour associated factors and their effect on uPAR cleavage, and the potential implications for cell proliferation, migration and invasion.MethodsMouse uPAR was stably overexpressed in the mouse OSCC cell line AT84. The ratio of full-length versus cleaved uPAR as analysed by Western blotting and its regulation was assessed by addition of different protease inhibitors and transforming growth factor - β1 (TGF-β1). The role of uPAR cleavage in cell proliferation and migration was analysed using real-time cell analysis and invasion was assessed using the myoma invasion model.ResultsWe found that when uPAR was overexpressed a proportion of the receptor was cleaved, thus the cells presented both full-length uPAR and uPAR (II-III). Cleavage was mainly performed by serine proteases and urokinase plasminogen activator (uPA) in particular. When the OSCC cells were stimulated with TGF-β1, the production of the uPA inhibitor PAI-1 was increased, resulting in a reduction of uPAR cleavage. By inhibiting cleavage of uPAR, cell migration was reduced, and by inhibiting uPA activity, invasion was reduced. We could also show that medium containing soluble uPAR (suPAR), and cleaved soluble uPAR (suPAR (II-III)), induced migration in OSCC cells with low endogenous levels of uPAR.ConclusionsThese results show that soluble factors in the tumour microenvironment, such as TGF-β1, PAI-1 and uPA, can influence the ratio of full length and uPAR (II-III) and thereby potentially effect cell migration and invasion. Resolving how uPAR cleavage is controlled is therefore vital for understanding how OSCC progresses and potentially provides new targets for therapy.

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Cellular glycosylphosphatidylinositol-specific phospholipase D regulates urokinase receptor shedding and cell surface expression

Cited by 127 publications

References 57 publications

Plasmin cleaves the juxtamembrane domain and releases truncated species of the urokinase receptor (CD87) from human bronchial epithelial cells

Plasmin cleaves the juxtamembrane domain and releases truncated species of the urokinase receptor (CD87) from human bronchial epithelial cells

Characterization of the five novel Ly‐6 superfamily members encoded in the MHC, and detection of cells expressing their potential ligands

Cleavage of the urokinase receptor (uPAR) on oral cancer cells: regulation by transforming growth factor – β1 (TGF-β1) and potential effects on migration and invasion

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