 |
PDBsum entry 5lhn
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Hydrolase/antibody
|
PDB id
|
|
|
|
5lhn
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Discovery of a novel conformational equilibrium in urokinase-Type plasminogen activator.
|
|
|
Authors
|
|
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.
|
|
|
Ref.
|
|
Sci Rep, 2017,
7,
3385.
|
|
|
PubMed id
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
