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TATIONpRÆSEN AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Danish-Chinese Centre for Proteases and Cancer The Danish National.

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Presentation on theme: "TATIONpRÆSEN AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Danish-Chinese Centre for Proteases and Cancer The Danish National."— Presentation transcript:

1 TATIONpRÆSEN AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Danish-Chinese Centre for Proteases and Cancer The Danish National Research Foundation The National Natural Science Foundation of China Allosteric Inhibition of Murine Urokinase-Type Plasminogen Activator The intradomain linker prevents allosteric transition of murine urokinase-type plasminogen activator into an inactive zymogen-like conformation with a blocked active-site Tobias Kromann-Hansen & Hans Peter Sørensen

2 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Background ›mU1 is a monoclonal antibody selected against murine uPA by the Finsen Laboratory at Rigshospitalet (Lund, Hansen et al. JBC 2008) ›mU1 successfully targets uPA dependent processes in vivo 2 Lund, Hansen et al. JBC 2008

3 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Activation of pro-uPA 3 ›uPA activation occurs through a single peptide bond cleavage at Lys15-Ile16 ›The newly formed N-terminus is inserted into a hydrophobic pocket and forms a salt-bridge to Asp194 ›This interaction stabilizes the active-site Andreasen, Kjøller et al., Int. J. Cancer, 1997

4 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark mU1 is not binding directly to the active site - mU1 is an allosteric antibody competitively inhibiting murine uPA - Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation

5 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark mU1 is not binding directly to the active site - mU1 is an allosteric antibody competitively inhibiting murine uPA - 5

6 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark mU1 inhibits the catalytic domain 20-fold more potently than the full length enzyme 6 Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation

7 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark The linker region has functional interactions to the catalytic domain 7 Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation

8 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark The linker region has functional interactions to the catalytic domain 8 Kratz, Mackman et al. Chem. Biol., 2000

9 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark Binding of mU1 to murine uPA exposes the N-terminus 9 Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation

10 Allosteric Inhibition of Murine uPA Tobias Kromann-Hansen Ph.d. Stud. AARHUS UNIVERSITY Department of Molecular Biology and Genetics Denmark The intradomain linker prevents allosteric transition of murine urokinase- type plasminogen activator into an inactive zymogen-like conformation with a blocked active-site 10

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