PDBsum entry 5lhs

Hydrolase

PDB id: 5lhs

Contents

Protein chains

224 a.a.

Ligands

SO4 ×8

Metals

_NI ×4

PDB id:

5lhs

Name:

Hydrolase

Title:

The ligand free catalytic domain of murine urokinase-type plasminogen activator

Structure:

Urokinase-type plasminogen activator. Chain: b, a, c, d. Synonym: upa. Engineered: yes. Mutation: yes

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Gene: plau. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

3.05Å R-factor: 0.216 R-free: 0.260

Authors:

T.Kromann-Hansen,E.L.Lange,H.P.Sorensen,G.H.Ghassabeh,M.Huang, J.K.Jensen,S.Muyldermans,P.J.Declerck,P.A.Andreasen

Key ref:

T.Kromann-Hansen et al. (2017). Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator. Sci Rep, 7, 3385. PubMed id: 28611361

Date:

12-Jul-16 Release date: 28-Jun-17

Protein chains

P06869  (UROK_MOUSE) -  Urokinase-type plasminogen activator from Mus musculus

Seq: 433 a.a.

Struc: 224 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.3.4.21.73 - u-plasminogen activator.
• Reaction: Specific cleavage of Arg-|-Val bond in plasminogen to form plasmin.

Sci Rep 7:3385 (2017)

PubMed id: 28611361

Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator.

T.Kromann-Hansen, E.Louise Lange, H.Peter Sørensen, G.Hassanzadeh-Ghassabeh, M.Huang, J.K.Jensen, S.Muyldermans, P.J.Declerck, E.A.Komives, P.A.Andreasen.

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.