5LHP

The p-aminobenzamidine active site inhibited catalytic domain of murine urokinase-type plasminogen activator in complex with the allosteric inhibitory nanobody Nb7

Summary for 5LHP

• Entry DOI: 10.2210/pdb5lhp/pdb
• Descriptor: Urokinase-type plasminogen activator, Camelid-Derived Antibody Fragment, SULFATE ION, ... (6 entities in total)
• Functional Keywords: trypsin-like serine proteases, nanobody, inhibitor, hydrolase
• Biological source: Mus musculus (Mouse); More
• Cellular location: Secreted: P06869
• Total number of polymer chains: 2
• Total formula weight: 45463.28

Authors

Kromann-Hansen, T.,Lange, E.L.,Sorensen, H.P.,Ghassabeh, G.H.,Huang, M.,Jensen, J.K.,Muyldermans, S.,Declerck, P.J.,Andreasen, P.A. (deposition date: 2016-07-12, release date: 2017-06-28, Last modification date: 2024-01-10)

Primary citation

Kromann-Hansen, T.,Louise Lange, E.,Peter Srensen, H.,Hassanzadeh-Ghassabeh, G.,Huang, M.,Jensen, J.K.,Muyldermans, S.,Declerck, P.J.,Komives, E.A.,Andreasen, P.A.
Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator.
Sci Rep, 7:3385-3385, 2017

PubMed Abstract: Although trypsin-like serine proteases have flexible surface-exposed loops and are known to adopt higher and lower activity conformations, structural determinants for the different conformations have remained largely obscure. The trypsin-like serine protease, urokinase-type plasminogen activator (uPA), is central in tissue remodeling processes and also strongly implicated in tumor metastasis. We solved five X-ray crystal structures of murine uPA (muPA) in the absence and presence of allosteric molecules and/or substrate-like molecules. The structure of unbound muPA revealed an unsuspected non-chymotrypsin-like protease conformation in which two β-strands in the core of the protease domain undergoes a major antiparallel-to-parallel conformational transition. We next isolated two anti-muPA nanobodies; an active-site binding nanobody and an allosteric nanobody. Crystal structures of the muPA:nanobody complexes and hydrogen-deuterium exchange mass spectrometry revealed molecular insights about molecular factors controlling the antiparallel-to-parallel equilibrium in muPA. Together with muPA activity assays, the data provide valuable insights into regulatory mechanisms and conformational flexibility of uPA and trypsin-like serine proteases in general.

PubMed: 28611361
DOI: 10.1038/s41598-017-03457-7

Experimental method

X-RAY DIFFRACTION (2.63 Å)