Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis.
Ann Louring Roldan,Maria Vittoria Cubellis,Maria Teresa Masucci,Niels Behrendt,Leif R. Lund,Keld Danø,Ettore Appella,Francesco Blasi +7 more
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Caseinolytic plaque assay, immunofluorescence analysis, direct binding studies and cross‐linking experiments show that the transfected mouse LB6 cells specifically bind human uPA, which in turn activates plasminogen.Abstract:
The surface receptor for urokinase plasminogen activator (uPAR) has been recognized in recent years as a key molecule in regulating plasminogen mediated extracellular proteolysis. Surface plasminogen activation controls the connections between cells, basement membrane and extracellular matrix, and therefore the capacity of cells to migrate and invade neighboring tissues. We have isolated a 1.4 kb cDNA clone coding for the entire human uPAR. An oligonucleotide synthesized on the basis of the N-terminal sequence of the purified protein was used to screen a cDNA library made from SV40 transformed human fibroblasts [Okayama and Berg (1983) Mol. Cell Biol., 3, 280-289]. The cDNA encodes a protein of 313 amino acids, preceded by a 21 residue signal peptide. A hydrophobicity plot suggests the presence of a membrane spanning domain close to the C-terminus. The cDNA hybridizes to a 1.4 kb mRNA from human cells, a size very close to that of the cloned cDNA. Expression of the uPAR cDNA in mouse cells confirms that the clone is complete and expresses a functional uPA binding protein, located on the cell surface and with properties similar to the human uPAR. Caseinolytic plaque assay, immunofluorescence analysis, direct binding studies and cross-linking experiments show that the transfected mouse LB6 cells specifically bind human uPA, which in turn activates plasminogen. The Mr of the mature human receptor expressed in mouse cells is approximately 55,000, in accordance with the naturally occurring, highly glycosylated human uPAR. The Mr calculated on the basis of the cDNA sequence, approximately 35,000, agrees well with that of the deglycosylated receptor.read more
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Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product
Donald P. Bottaro,Jeffrey S. Rubin,D. L. Faletto,Andrew K Chan,Thomas E. Kmiecik,G F Vande Woude,Stuart A. Aaronson +6 more
TL;DR: A 145-kilodalton tyrosyl phosphoprotein observed in rapid response to HGF treatment of intact target cells was identified by immunoblot analysis as the beta subunit of the c-met proto-oncogene product, a membrane-spanning tyrosine kinase.
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Mechanism Of Cell Surface Activation Of 72-kDa Type IV Collagenase ISOLATION OF THE ACTIVATED FORM OF THE MEMBRANE METALLOPROTEASE
Alex Y. Strongin,Ivan E. Collier,Gregory A. Bannikov,Barry L. Marmer,Gregory A. Grant,Gregory I. Goldberg +5 more
TL;DR: Activation of 72T4Cl on the cell membrane provides a basic mechanism for spatially regulated extracellular proteolysis and presents a new target for prognosis and treatment of metastatic disease.
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The urokinase-type plasminogen activator system in cancer metastasis: a review
TL;DR: Recent observations related to the molecular and cellular mechanisms underlying the role of the u‐PA system are discussed, suggesting that the system does not support tumor metastasis by the unrestricted enzyme activity of u‐ PA and plasmin and that pericellular molecular and functional interactions appear to allow temporal and spatial re‐organizations of the system during cell migration.
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The plasminogen activation system in tumor growth, invasion, and metastasis.
TL;DR: The increased knowledge about the plasminogen activation system may allow utilization of its components as targets for anti-invasive therapy.
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Biochemistry and Molecular Biology of Gelatinase B or Matrix Metalloproteinase-9 (MMP-9)
Philippe E. Van den Steen,Bénédicte Dubois,Inge Nelissen,Pauline M. Rudd,Raymond A. Dwek,Ghislain Opdenakker +5 more
TL;DR: The ability of gelatinase B to degrade components of the extracellular matrix and to regulate the activity of a number of soluble proteins confers an important role in various physiological and pathological processes, including reproduction, growth, development, inflammation, and vascular and proliferative diseases.
References
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Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Book ChapterDOI
Plasminogen activators, tissue degradation, and cancer.
Keld Danø,Peter A. Andreasen,J Grøndahl-Hansen,Peter Marcus Kristensen,L.S. Nielsen,L. Skriver +5 more
TL;DR: This chapter describes two types of plasminogen activators—namely, the urokinase-type plasMinogen activator (u-PA) and the tissue- type plasmineg activator(t-PA), which are essentially different gene products.
Journal ArticleDOI
Monoclonal antibodies to human 54,000 molecular weight plasminogen activator inhibitor from fibrosarcoma cells--inhibitor neutralization and one-step affinity purification.
L.S. Nielsen,Peter A. Andreasen,J Grøndahl-Hansen,Jian-Ying Huang,Peter Marcus Kristensen,Keld Danø +5 more
TL;DR: Mouse monoclonal antibodies derived against a plasminogen activator inhibitor derived from cultured human umbilical cord endothelial cells identified four clones of hybridomas producing IgG1 antibodies which could be used for immunocytochemical localization of the inhibitor in HT-1080 cells.