Forskning
Udskriv Udskriv
Switch language
Region Hovedstaden - en del af Københavns Universitetshospital
Udgivet

Stabilizing a flexible interdomain hinge region harboring the SMB binding site drives uPAR into its closed conformation

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

DOI

  1. Lactate-Mediated Protection of Retinal Ganglion Cells

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  2. The Limitations of In Vitro Experimentation in Understanding Biofilms and Chronic Infection

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  3. Crystal Structure of the Urokinase Receptor in a Ligand-Free Form

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  4. Multiple barriers to recombination between divergent HIV-1 variants revealed by a dual-marker recombination assay

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  5. Quorum sensing regulation in Aeromonas hydrophila

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  1. ANGPTL4 inactivates lipoprotein lipase by catalyzing the irreversible unfolding of LPL's hydrolase domain

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  2. Intermittent chylomicronemia caused by intermittent GPIHBP1 autoantibodies

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  3. Unfolding of monomeric lipoprotein lipase by ANGPTL4: Insight into the regulation of plasma triglyceride metabolism

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  4. Crystal Structures of Human C4.4A Reveal the Unique Association of Ly6/uPAR/α-neurotoxin Domain

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Vis graf over relationer

The urokinase-type plasminogen activator receptor (uPAR) is a multidomain glycolipid-anchored membrane protein, which facilitates extracellular matrix remodeling by focalizing plasminogen activation to cell surfaces via its high-affinity interaction with uPA. The modular assembly of its three LU domains is inherently flexible and binding of uPA drives uPAR into its closed conformation, which presents the higher affinity state for vitronectin thus providing an allosteric regulatory mechanism. Using a new class of epitope mapped monoclonal anti-uPAR antibodies, we now demonstrate that the reciprocal stabilization is indeed also possible. By surface plasmon resonance studies we show that these mAbs and vitronectin have overlapping binding sites on uPAR and that they share Arg(91) as hotspot residue in their binding interfaces. The crystal structure solved for one of these uPAR•mAb complexes at 3.0Å clearly shows that this mAb preselects the closed uPAR conformation with an empty, but correctly assembled large hydrophobic binding cavity for uPA. Accordingly, these mAbs inhibit the uPAR-dependent lamellipodia formation and migration on vitronectin coated matrices irrespective of the conformational status of uPAR and its occupancy with uPA. This is to the best of our knowledge the first study, showing that the dynamic assembly of the three LU domains in uPAR(wt) can be driven towards the closed form by an external ligand, which is not engaging the hydrophobic uPA-binding cavity. As this binding interface is also exploited by the SMB domain of vitronectin this relationship should therefore be taken into consideration when exploring uPAR-dependent cell adhesion and migration in vitronectin-rich environments.

OriginalsprogEngelsk
TidsskriftJournal of Molecular Biology
Vol/bind427
Sider (fra-til)1389-1403
ISSN0022-2836
DOI
StatusUdgivet - 2015

ID: 44975277