TATION pRÆSEN 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
Transcript
TATIONpRÆSEN
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
Danish-Chinese Centre for Proteases and CancerThe 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
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
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
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Lund, Hansen et al. JBC 2008
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
Activation of pro-uPA
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› 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-siteAndreasen, Kjøller et al., Int. J. Cancer, 1997
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
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
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
mU1 is not binding directly to the active site- mU1 is an allosteric antibody competitively inhibiting murine uPA -
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Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
mU1 inhibits the catalytic domain 20-fold more potently than the full length enzyme
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Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
The linker region has functional interactions to the catalytic domain
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Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
The linker region has functional interactions to the catalytic domain
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Kratz , Mackman et al. Chem. Biol., 2000
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
Binding of mU1 to murine uPA exposes the N-terminus
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Kromann-Hansen, T., Lund, IK., Zhou, L. Andreasen, PA., Høyer-Hansen, G., Sørensen, HP. In preparation
Allosteric Inhibition of Murine uPATobias Kromann-Hansen
Ph.d. Stud.
AARHUS UNIVERSITYDepartment of Molecular Biology and GeneticsDenmark
The intradomain linker prevents allosteric transition of murine urokinase-type plasminogen activator into an inactive zymogen-like conformation with a blocked active-site
