Evolution and Medical Significance of LU Domain−Containing Proteins

Leth, Julie Maja; Leth-Espensen, Katrine Zinck; Kristensen, Kristian Kølby; Kumari, Anni; Winther, Anne-Marie Lund; Young, Stephen G.; Ploug, Michael

doi:10.3390/ijms20112760

Cited by 30 publications

(36 citation statements)

References 121 publications

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“…Inflammation and oxidative stress are central components of the pathogenesis of acute kidney injury, implicating multiple subtypes of immune cells. 8,9 Evidence of a pathway linking the bone marrow to kidney injury has emerged, involving soluble urokinase plasminogen activator receptor (suPAR) 7,[10][11][12][13][14][15][16][17] -the circulating form of a glycosylphosphatidylinositol-anchored three-domain membrane protein. This receptor is normally expressed at very low levels on a variety of cells, including endothelial cells, podocytes, and, with induced expression, immunologically active cells such as monocytes and lymphocytes.…”

Section: Results-mentioning

confidence: 99%

Soluble Urokinase Receptor and Acute Kidney Injury

et al. 2020

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BACKGROUND-Acute kidney injury is common, with a major effect on morbidity and health care utilization. Soluble urokinase plasminogen activator receptor (suPAR) is a signaling glycoprotein thought to be involved in the pathogenesis of kidney disease. We investigated whether a high level of suPAR predisposed patients to acute kidney injury in multiple clinical contexts, and we used experimental models to identify mechanisms by which suPAR acts and to assess it as a therapeutic target. METHODS-We measured plasma levels of suPAR preprocedurally in patients who underwent coronary angiography and patients who underwent cardiac surgery and at the time of admission to the intensive care unit in critically ill patients. We assessed the risk of acute kidney injury at 7 days as the primary outcome and acute kidney injury or death at 90 days as a secondary outcome, according to quartile of suPAR level. In experimental studies, we used a monoclonal antibody to urokinase plasminogen activator receptor (uPAR) as a therapeutic strategy to attenuate acute kidney injury in transgenic mice receiving contrast material. We also assessed cellular bioenergetics and generation of reactive oxygen species in human kidney proximal tubular (HK-2) cells that were exposed to recombinant suPAR.

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Section: Results-mentioning

confidence: 99%

Soluble Urokinase Receptor and Acute Kidney Injury

et al. 2020

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“…Proteins with Ly6/uPAR/α-neurotoxin domain (LU-domain), also named three-fingered protein domain or TFPD, are widespread in the animal kingdom and mainly comprises either secreted or glycosyl-phosphatidylinositol (GPI) anchored single domain proteins with diverse biological functions [1]. The hallmark of a prototypical LU-domain is 8 conserved cysteine residues engaged in a defined disulfide-bonding, which forms a compact cysteine-rich knot (palm) projecting three extended loops (fingers) stabilized by 5-6 β-strands [2][3][4].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2020

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Ly6/uPAR/α-neurotoxin domain (LU-domain) is characterized by the presence of 4-5 disulfide bonds and three flexible loops that extend from a core stacked by several conversed disulfide bonds (thus also named three-fingered protein domain). This highly structurally stable protein domain is typically a protein-binder at extracellular space. Most LU proteins contain only single LU-domain as represented by Ly6 proteins in immunology and α-neurotoxins in snake venom. For Ly6 proteins, many are expressed in specific cell lineages and in differentiation stages, and are used as markers. In this study, we report the crystal structures of the two LU-domains of human C4.4A alone and its complex with a Fab fragment of a monoclonal anti-C4.4A antibody. Interestingly, both structures showed that C4.4A forms a very compact globule with two LU-domain packed face to face. This is in contrast to the flexible nature of most LU-domain-containing proteins in mammals. The Fab combining site of C4.4A involves both LU-domains, and appears to be the binding site for AGR2, a reported ligand of C4.4A. This work reports the first structure that contain two LU-domains and provides insights on how LU-domains fold into a compact protein and interacts with ligands.

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“…The urokinase‐type plasminogen activator receptor (uPAR) is a glycophosphatidylinositol (GPI)‐anchored cell surface receptor belonging to the LU‐protein domain family [1,2] . Although uPAR is reported to be at the center of a complex network of protein‐protein interactions, its immediate binding partners are the serine proteinase urokinase‐type plasminogen activator uPA [3] and the extracellular matrix protein vitronectin [4–6] .…”

Section: Introductionmentioning

confidence: 99%

“…The urokinase-type plasminogen activator receptor (uPAR) is a glycophosphatidylinositol (GPI)-anchored cell surface receptor belonging to the LU-protein domain family. [1,2] Although uPAR is reported to be at the center of a complex network of proteinprotein interactions, its immediate binding partners are the serine proteinase urokinase-type plasminogen activator uPA [3] and the extracellular matrix protein vitronectin. [4][5][6] The interaction with uPA is responsible for uPAR's beneficial role in clearing extravascular fibrin deposits by plasmin-dependent fibrinolysis, [7] but particularly in the setting of chronic inflammatory conditions it does also have deleterious effects exacerbating the pathological of e. g. arthritis [8,9] and its expression is correlated to poor patient prognosis in many solid cancers allegedly by facilitating tumor invasion and metastasis.…”

Section: Introductionmentioning

confidence: 99%

Small‐Molecule Inhibition of the uPAR ⋅ uPA Interaction by Conformational Selection

Bum‐Erdene

Leth

et al. 2020

ChemMedChem

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The urokinase receptor (uPAR) is a cell surface receptor that binds to the serine protease urokinase‐type plasminogen activator (uPA) with high affinity. This interaction is beneficial for extravascular fibrin clearance, but it has also been associated with a broad range of pathological conditions including cancer, atherosclerosis, and kidney disease. Here, starting with a small molecule that we previously discovered by virtual screening and cheminformatics analysis, we design and synthesize several derivatives that were tested for binding and inhibition of the uPAR ⋅ uPA interaction. To confirm the binding site and establish a binding mode of the compounds, we carried out biophysical studies using uPAR mutants, among them uPARH47C−N259C, a mutant previously developed to mimic the structure of uPA‐bound uPAR. Remarkably, a substantial increase in potency is observed for inhibition of uPARH47C−N259C binding to uPA compared to wild‐type uPAR, consistent with our use of the structure of uPAR in its uPA‐bound state to design small‐molecule uPAR ⋅ uPA antagonists. Combined with the biophysical studies, molecular docking followed by extensive explicit‐solvent molecular dynamics simulations and MM‐GBSA free energy calculations yielded the most favorable binding pose of the compound. Collectively, these results suggest that potent inhibition of uPAR binding to uPA with small molecules will likely only be achieved by developing small molecules that exhibit high‐affinity to solution apo structures of uPAR, rather than uPA‐bound structures of the receptor.

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Evolution and Medical Significance of LU Domain−Containing Proteins

Cited by 30 publications

References 121 publications

Soluble Urokinase Receptor and Acute Kidney Injury

Soluble Urokinase Receptor and Acute Kidney Injury

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

Small‐Molecule Inhibition of the uPAR ⋅ uPA Interaction by Conformational Selection

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