title: Inhibition of the interaction of urokinase-type plasminogen activator (uPA) with its receptor (uPAR) by synthetic peptides.
abstract: Focusing of the serine protease urokinase-type plasminogen activator (uPA) to the cell surface via interaction with its specific receptor (uPAR, CD87) is an important step for tumor cell invasion and metastasis. The ability of a synthetic peptide derived from the uPAR-binding region of uPA (comprising amino acids 16-32 of uPA; uPA(16-32)) to inhibit binding of fluorescently labeled uPA to uPAR on human promyeloid U937 cells was assessed by quantitative flow cytofluorometric analysis (FACS) and compared to the inhibitory capacities of other synthetic peptides known to interfere with uPA/uPAR-interaction. An about 3000-fold molar excess of uPA(16-32) resulted in 50% inhibition of pro-uPA binding to cell surface-associated uPAR. Using a solid-phase uPA-ligand binding assay employing recombinant soluble uPAR coated to microtiter plates, the minimal binding region of wild-type uPA was determined. The linear peptide uPA(19-31) and its more stable disulfide-bridged cyclic form (cyclo(19,31)uPA(19-31)) displayed uPAR-binding activity whereas other peptides such as uPA(18-30), uPA(20-32) or uPA(20-30) did not react with uPAR. Cyclic peptide derivatives of cyclo(19,31)uPA(19-31) in which certain amino acids were deleted and/or replaced by other amino acids as well as uPAR-derived wild-type peptides did also not inhibit uPA/uPAR-interaction. Therefore, the present investigations identified cyclo(19,31)uPA(19-31) as a potential lead structure for the development of uPA-peptide analogues to block uPA/uPAR-interaction.
authors.affiliation: Institut für Organische Chemie und Biochemie der Technischen Universität Munchen, Garching, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Bürgle
authors.forename: M
authors.initials: M
authors.lastname: Koppitz
authors.forename: C
authors.initials: C
authors.lastname: Riemer
authors.forename: H
authors.initials: H
authors.lastname: Kessler
authors.forename: B
authors.initials: B
authors.lastname: König
authors.forename: U H
authors.initials: UH
authors.lastname: Weidle
authors.forename: J
authors.initials: J
authors.lastname: Kellermann
authors.forename: F
authors.initials: F
authors.lastname: Lottspeich
authors.forename: H
authors.initials: H
authors.lastname: Graeff
authors.forename: M
authors.initials: M
authors.lastname: Schmitt
authors.forename: L
authors.initials: L
authors.lastname: Goretzki
authors.forename: U
authors.initials: U
authors.lastname: Reuning
authors.forename: O
authors.initials: O
authors.lastname: Wilhelm
authors.forename: V
authors.initials: V
authors.lastname: Magdolen
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D005434:Flow Cytometry; D006801:Humans; D008024:Ligands; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D008024:Ligands; C524898:PLAUR protein, human; D010455:Peptides; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator