A5037566190- Protease and Inhibitor Mechanisms
- Blood Coagulation and Thrombosis Mechanisms
- Peptidase Inhibition and Analysis
- Cell Adhesion Molecules Research
- Monoclonal and Polyclonal Antibodies Research
- Signaling Pathways in Disease
- S100 Proteins and Annexins
- Enzyme Production and Characterization
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Complement system in diseases
- Nanoparticle-Based Drug Delivery
- Connective tissue disorders research
- HER2/EGFR in Cancer Research
- Lymphatic System and Diseases
- Platelet Disorders and Treatments
- Hemophilia Treatment and Research
- Blood properties and coagulation
- Bone and Dental Protein Studies
- Cellular transport and secretion
- Erythrocyte Function and Pathophysiology
- Occupational and environmental lung diseases
- RNA Interference and Gene Delivery
- Immune cells in cancer
- Bone Metabolism and Diseases
- Connective Tissue Growth Factor Research
Rigshospitalet
2015-2025
University of Copenhagen
2015-2025
Technical University of Denmark
2023
Centre de Nanosciences et de Nanotechnologies
2017
Copenhagen University Hospital
2004-2014
Kyowa Kirin (France)
2013
Business Innovation Centre
2011
Capital Region of Denmark
1988-2006
National Institute of Dental and Craniofacial Research
1999
National Institutes of Health
1999
The cellular receptor for human urokinase-type plasminogen activator (u-PAR) is shown by several independent criteria to be a true member of family integral membrane proteins, anchored the plasma exclusively COOH-terminal glycosyl-phosphatidylinositol moiety. 1) Amino acid analysis u-PAR after micropurification affinity chromatography and N-[2-hydroxy-1,1-bis(hydroxymethyl)-ethyl]glycine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed presence 2-3 mol ethanolamine/mol...
The specific cellular receptor for urokinase-type plasminogen activator (uPA) is found on a variety of cell types and has been postulated to play central role in the mediation pericellular proteolytic activity. We have studied kinetics (Plg) activation catalyzed by uPA specifically bound its human monocytoid cell-line U937 demonstrate this process properties differing widely from those observed solution. solution-phase reaction was characterized Km 25 microM cell-associated fell 40-fold 0.67...
Tissue remodeling processes critically depend on the timely removal and of preexisting collagen scaffolds. Nevertheless, many aspects related to turnover this abundant extracellular matrix component in vivo are still incompletely understood. We therefore took advantage recent advances optical imaging develop an assay visualize situ identify cell types molecules involved process. Collagen introduced into dermis mice underwent cellular endocytosis a partially metalloproteinase–dependent manner...
Human HT-1080 fibrosarcoma cells produce urokinase-type plasminogen activator (u-PA) and type 1 inhibitor (PAI-1). We found that after incubation of monolayer cultures with purified native human in serum-containing medium, bound plasmin activity could be eluted from the tranexamic acid, an analogue lysine. The was result activation on cell surface; not taken up onto deliberate addition to medium. surface formation inhibited by anticatalytic monoclonal antibody u-PA, indicating this enzyme...
The receptor for human urokinase-type plasminogen activator (u-PA) was purified from phorbol 12-myristate 13-acetate-stimulated U937 cells by temperature-induced phase separation of detergent extracts, followed affinity chromatography with immobilized diisopropyl fluorophosphate-treated u-PA. protein shows a single 55-60 kDa band after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. It is heavily glycosylated protein, the deglycosylated polypeptide chain...
The iodinated Mr approximately equal to 15,000 amino-terminal fragment (ATF) of the urokinase-type plasminogen activator (u-PA) molecule bound specifically cell surface all seven cultured human tumor lines studied. Cross-linking ATF using a bifunctional amino-reactive reagent followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed with four studied occurrence single band migrating an 70,000-75,000, indicating complex formation 55,000-60,000 u-PA...
Urokinase-type plasminogen activator (uPA) binds to a specific receptor on various cell types, the bound molecule retaining its enzymatic activity against plasminogen. We have now investigated whether receptor-bound uPA also retains ability react with and be inhibited by inhibitors (PAI-1 PAI-2). human U937 monocyte-like cells was PAI-1 (in active form in presence of vitronectin fragments) an association rate constant 4.5 x 10(6) M-1 s-1, which 40% lower than that obtained for solution (7.9...
The purified urokinase plasminogen activator receptor (u-PAR) was cleaved into two fragments by mild chymotrypsin treatment. smaller fragment (apparent Mr 16,000) possessed the ligand-binding capability, as shown chemical cross-linking analysis. This constituted NH2-terminal part of intact receptor, probably including whole sequence 1-87, and contained N-linked carbohydrate. After detergent phase separation in Triton X-114 system, present water where its binding activity could be...
The cellular receptor for urokinase-type plasminogen activator (uPAR) is a glycolipid-anchored three-domain membrane protein playing central role in pericellular activation. We have found that urokinase (uPA) can cleave its between domains 1 and 2 generating cell-associated uPAR variant without ligand-binding properties. In extracts of U937 cells there are two variants which after complete deglycosylation apparent molecular masses 35,000 27,000. Analysis with monoclonal antibodies showed...
We have raised four monoclonal antibodies recognizing different epitopes within the human cell‐surface receptor for urokinase‐type plasminogen activator (u‐PA). One of these completely abolishes potentiation plasmin generation observed upon incubation zymogens pro‐u‐PA and with U937 cells. This antibody, which is also only one to inhibit binding DFP‐inactivated [ 125 I]‐u‐PA cells, directed against u‐PA NH 2 ‐terminal domain u‐PAR, a well‐defined fragment formed by limited chymotrypsin...
Physiologic turnover of interstitial collagen is mediated by a sequential pathway in which fragmented pericellular collagenases, endocytosed receptors, and routed to lysosomes for degradation cathepsins. Here, we use intravital microscopy investigate if malignant tumors, are characterized high rates extracellular matrix turnover, utilize similar pathway. Tumors epithelial, mesenchymal, or neural crest origin all display vigorous endocytic degradation. The cells engaged this process...
The uptake and lysosomal degradation of collagen by fibroblasts constitute a major pathway in the turnover connective tissue. However, molecular mechanisms governing this are poorly understood. Here, we show that urokinase plasminogen activator receptor–associated protein (uPARAP)/Endo180, novel mesenchymally expressed member macrophage mannose receptor family endocytic receptors, is key player process. Fibroblasts from mice with targeted deletion uPARAP/Endo180 gene displayed near to...
The urokinase receptor (uPAR) is a for both plasminogen activator (uPA) and the adhesion protein vitronectin. There are two forms of cell surface-bound uPAR; intact uPAR cleaved form, uPAR(2+3), which formed by uPA-catalyzed cleavage uPAR. In ligand-blotting experiments we found that vitronectin binds but not uPAR(2+3). real-time biomolecular interaction analysis using recombinant, soluble (suPAR) plasma multimeric bound to intact, antibody-immobilized suPAR. Monoclonal antibodies against...
The plasminogen activation cascade system, directed by urokinase and the receptor, plays a key role in extracellular proteolysis during tissue remodeling. To identify molecular interaction partners of these trigger proteins on cell, we combined covalent protein cross-linking with mass spectrometry based methods for peptide mapping primary structure analysis electrophoretically isolated conjugates. A specific tri-molecular complex was observed upon addition pro-urokinase to human U937 cells....
A specific surface receptor for urokinase plasminogen activator (uPA) recognizes the amino-terminal growth factor-like sequence of uPA, a region independent from and not required catalytic activity this enzyme. The properties uPA (uPAR) localization distribution in tumor cells tissues suggest that uPA/uPAR interaction may be important regulating extracellular proteolysis-dependent processes (e.g., invasion, tissue destruction). Phorbol myristate acetate (PMA), an inducer U937 cell...
The collagens of the extracellular matrix are most abundant structural proteins in mammalian body. In tissue remodeling and invasive growth malignant tumors, constitute an important barrier, consequently, turnover collagen is a rate-limiting process these events. A recently discovered route with importance for tumor involves intracellular degradation governed by receptor, urokinase plasminogen activator receptor-associated protein (uPARAP or Endo180). interplay between this mechanism...
We recently reported that uPARAP/Endo180 can mediate the cellular uptake and lysosomal degradation of collagen by cultured fibroblasts. Here, we show has a key role in during mammary carcinoma progression. In normal murine gland, is widely expressed periductal fibroblast-like mesenchymal cells line epithelial cells. This pattern expression preserved polyomavirus middle T–induced carcinogenesis, with strong embedded within collagenous stroma surrounding nests uPARAP/Endo180-negative tumor...
Processes such as cell proliferation, angiogenesis, apoptosis, or invasion are strongly influenced by the surrounding microenvironment of tumor. Therefore, ability to change these surroundings represents an important property through which tumor cells able acquire specific functions necessary for growth and dissemination. Matrix metalloproteinases (MMPs) constitute key players in this process, allowing modify extracellular matrix (ECM) release cytokines, factors, other cell-surface...