| 2007 |
LIMP-2 is a specific binding partner of β-glucocerebrosidase (GCase) and serves as the mannose-6-phosphate-independent lysosomal targeting receptor for GCase. The interaction involves a coiled-coil domain within the lumenal domain of LIMP-2. In LIMP-2-deficient mice, GCase is secreted rather than delivered to lysosomes, and reconstitution of LIMP-2 rescues lysosomal GCase levels and distribution. |
Affinity chromatography, LIMP-2-deficient mouse fibroblasts and macrophages, reconstitution experiments, subcellular fractionation |
Cell |
High |
18022370
|
| 1998 |
AP-3 selectively binds the cytoplasmic tail of LIMP-2 via a DEXXXLI dileucine-based motif, mediating sorting of LIMP-2 to lysosomes. AP-1 and AP-2 do not interact with this tail, establishing AP-3 as the specific adaptor for LIMP-2 lysosomal targeting. |
Surface plasmon resonance binding assay with recombinant AP complexes and cytoplasmic tail peptides |
The EMBO journal |
High |
9482728
|
| 2003 |
The [DE]XXXL[LI]-type dileucine signal in LIMP-2's cytoplasmic tail interacts specifically with the gamma-sigma1 subunits of AP-1 and delta-sigma3 subunits of AP-3, but not AP-2 or AP-4 hemicomplexes, defining the molecular basis of AP-3-mediated endosomal/lysosomal sorting. |
Yeast three-hybrid assay, in vitro binding to whole AP complexes |
The Journal of cell biology |
High |
14691137
|
| 2013 |
Crystal structure of LIMP-2 reveals a helical bundle where GCase binds and a large cavity/tunnel traversing the entire molecule, consistent with a lipid transport function. Mutagenesis of the tunnel in the SR-BI homologue indicates this cavity mediates cholesterol(ester) transfer from bound lipoproteins to the membrane. |
X-ray crystallography, homology modelling, site-directed mutagenesis of tunnel residues in SR-BI |
Nature |
High |
24162852
|
| 2009 |
Disease-causing AMRF nonsense mutations in LIMP-2 (W146SfsX16, W178X) abolish GCase binding, while the Q288X truncation retains near-normal binding. The missense mutation H363N increases GCase binding affinity. A coiled-coil domain (residues 145–288) is essential for GCase binding; disruption of the helical/amphipathic coiled-coil structure abolishes this interaction. |
Co-immunoprecipitation, binding assays with mutant LIMP-2 constructs, synthetic peptide studies |
Human molecular genetics |
High |
19933215
|
| 2012 |
A single histidine residue in LIMP-2 (H363) functions as a pH sensor required for GCase binding at neutral pH and its release in late endosomal/lysosomal acidic compartments. Vacuolar H+-ATPase-mediated lumenal acidification triggers dissociation of the LIMP-2/GCase complex. |
Site-directed mutagenesis of H363, pharmacological inhibition of V-ATPase, co-immunoprecipitation at different pH values |
Traffic (Copenhagen, Denmark) |
High |
22537104
|
| 2014 |
Structural analysis of LIMP-2 reveals that GCase binding and pH-dependent release is governed by a histidine trigger, and that LIMP-2 carries P-Man9GlcNAc2 at N325 enabling it to bind the mannose-6-phosphate receptor (MPR) with affinity similar to the LIMP-2/GCase interaction; β-GCase and MPR binding sites are functionally separate, allowing formation of a stable ternary LIMP-2/GCase/MPR complex demonstrated in living cells by FLIM. |
X-ray crystallography, surface plasmon resonance, fluorescence lifetime imaging microscopy (FLIM) in living cells |
Nature communications |
High |
25027712
|
| 2015 |
LIMP-2 lysosomal sorting occurs via a mannose-6-phosphate-independent pathway: in fibroblasts lacking MPRs or GlcNAc-1-phosphotransferase (the M6P-forming enzyme), LIMP-2 still localizes to lysosomes, and lysosomal LIMP-2 levels are comparable in wild-type and phosphotransferase-defective mouse liver. The M6P modification on the LIMP-2 ectodomain is dispensable for its lysosomal targeting. |
Immunofluorescence in MPR-deficient and GlcNAc-1-phosphotransferase-defective fibroblasts, lysosome purification and immunoblot from mouse liver, heterologous expression of LIMP-2 luminal domain |
Traffic (Copenhagen, Denmark) |
High |
26219725
|
| 2019 |
LIMP-2 mediates lysosomal cholesterol export: the luminal cavity can bind and deliver exogenous cholesterol to the lysosomal membrane and subsequently to lipid droplets. LIMP-2 depletion alters SREBP-2-mediated cholesterol regulation and LDL-receptor levels, and LIMP-2 operates in parallel with NPC proteins for lysosomal cholesterol export. |
Molecular modeling, crosslinking studies, microscale thermophoresis, cell-based cholesterol transport assays, SREBP-2/LDL-receptor immunoblotting in LIMP-2 knockout cells |
Nature communications |
High |
31387993
|
| 2000 |
The dileucine sorting motif in LIMP-2's C-terminal tail requires an acidic glutamate (Glu) at position -4 upstream of the critical leucine for efficient intracellular sorting to lysosomes, but this residue is dispensable for surface internalization by endocytosis, demonstrating distinct structural requirements for intracellular sorting versus endocytosis. |
Site-directed mutagenesis of LIMP-2 cytoplasmic tail, subcellular localization by microscopy, endocytosis assays in transfected cells |
The Journal of biological chemistry |
Medium |
10973972
|
| 1998 |
SR-BII (an alternatively spliced isoform of the SCARB2/SR-BI gene with a distinct C-terminal cytoplasmic tail) is enriched in caveolae and mediates both selective cellular uptake of cholesteryl ether from HDL and HDL-dependent cholesterol efflux, but with ~4-fold lower efficiency than SR-BI. |
Subcellular fractionation of CHO transfectants, radiolabeled HDL cholesteryl ether uptake assay, cholesterol efflux assay, adenoviral overexpression in vivo |
The Journal of biological chemistry |
High |
9614139
|
| 2004 |
SR-BII (the SCARB2 gene splice variant) is predominantly localized intracellularly (~80–90% intracellular vs ~70% surface for SR-BI) due to its distinct C-terminal cytoplasmic tail, and rapidly internalizes HDL via endocytosis, with internalized HDL co-localizing in the endosomal recycling compartment. Deletion of the SR-BI C-terminus does not affect its surface localization, confirming the SR-BII C-terminus confers intracellular targeting. |
Cell surface biotinylation, EGFP-tagged receptor imaging, pulse-chase HDL uptake experiments, subcellular co-localization with transferrin (endosomal recycling marker) |
The Journal of biological chemistry |
High |
14726519
|
| 2005 |
SR-BII mediates HDL endocytosis through a clathrin-dependent, caveolae-independent pathway. A dileucine motif at positions 492–493 in the SR-BII C-terminal cytoplasmic tail is required for HDL particle endocytosis; L492A substitution increases surface HDL binding and reduces endocytosis. Introduction of the SR-BII YTPLL motif into SR-BI converts it into an endocytic receptor. |
Site-directed mutagenesis of SR-BII tail residues, HDL endocytosis assays, clathrin/caveolin pathway inhibition, chimeric SR-BI/SR-BII constructs |
The Journal of biological chemistry |
High |
16368683
|
| 2012 |
SCARB2 functions as an uncoating receptor for EV71: after virus-SCARB2 complex internalization into endosomes, acidic pH (below 6.0) combined with SCARB2 triggers conversion of native EV71 virions into empty capsids lacking both genomic RNA and VP4. This uncoating does not occur with PSGL1 as receptor under any pH condition. |
Sucrose density gradient centrifugation analysis of viral uncoating, incubation of EV71 with L-SCARB2 cells or soluble SCARB2 at various pH values, immunofluorescence colocalization with endosomal markers |
Journal of virology |
High |
23302872
|
| 2014 |
Crystal structures of SCARB2 under neutral and acidic conditions reveal a pH-dependent conformational change that opens a lipid-transfer tunnel, enabling expulsion of a hydrophobic pocket factor from EV71, a prerequisite for viral uncoating. The canyon region of EV71 VP1 mediates receptor interaction, with key residues identified. |
X-ray crystallography of SCARB2 at neutral and acidic pH, structural comparison, mutagenesis of virus-receptor contact residues |
Protein & cell |
High |
24986489
|
| 2018 |
Cryo-EM structure of the EV71-SCARB2 complex at 3.4 Å resolution shows SCARB2 binds EV71 on the southern rim of the canyon (not across the canyon as predicted). Helices α5 (152–163) and α7 (183–193) of SCARB2 and the VP1 GH and VP2 EF loops of EV71 dominate the interaction, suggesting an allosteric mechanism for low-pH uncoating. |
Cryo-electron microscopy of EV71-SCARB2 complex |
Nature microbiology |
High |
30531980
|
| 2011 |
The region of human SCARB2 encompassing amino acids 142–204 is critical for EV71 virion binding and infection; chimeric SCARB2 constructs carrying this human region in a mouse Scarb2 backbone confer susceptibility to EV71, whereas those retaining the mouse sequence in this region do not support efficient viral binding. |
Human-mouse SCARB2 chimeric mutant expression in L929 cells, viral binding assays, infection assays |
Journal of virology |
High |
21389126
|
| 2012 |
EV71 entry into cells expressing SCARB2 occurs via a clathrin-mediated, pH-dependent, cholesterol-sensitive endocytic pathway; siRNA knockdown of clathrin or dynamin blocks entry, whereas caveolin knockdown does not affect entry. |
siRNA knockdown of clathrin, dynamin, caveolin; chemical inhibitors of clathrin-mediated endocytosis and caveolae-mediated endocytosis; pH perturbation experiments; cholesterol depletion |
PloS one |
Medium |
22272359
|
| 2015 |
SCARB2 mediates endosomal translocation of TLR9 in plasmacytoid dendritic cells (pDCs): SCARB2 knockdown results in retention of TLR9 in the ER and impaired nuclear translocation of IRF7, leading to dramatically reduced CpG-induced type I IFN production. SCARB2 localizes to late endosomes/lysosomes in pDCs and is required for TLR7-ligand-induced IFN-I as well. |
siRNA knockdown of SCARB2 in pDC cell line GEN2.2, subcellular localization by immunofluorescence, TLR9/IRF7 localization by immunofluorescence, IFN-I ELISA |
Journal of immunology |
Medium |
25862818
|
| 2014 |
The GCase-binding sequence on GCase for LIMP-2 is an 11-amino acid stretch (DSPIIVDITKD); Asp399, Ile402, and Ile403 are particularly important. Alanine substitution at any of these residues decreases GCase binding to LIMP-2, alters pH-dependent binding, diminishes lysosomal trafficking of GCase, and increases GCase secretion. The EV71-binding site on LIMP-2/SCARB2 is distinct from the GCase-binding site. |
Deletion constructs, alanine-scanning mutagenesis, binding assays, co-localization, GCase secretion assays |
The Journal of biological chemistry |
High |
25202012
|
| 2015 |
In human fibroblasts and neuron-like cells, GCase lysosomal targeting is completely dependent on LIMP-2, whereas in blood (lymphocytes), GCase is partially targeted to lysosomes by a LIMP-2-independent mechanism. Recombinant human GCase (enzyme replacement therapy) is taken up by cells independently of LIMP-2 but its lysosomal trafficking requires LIMP-2. |
GCase activity and localization in AMRF patient fibroblasts, lymphocytes, and neuronal model; LIMP-2-deficient and -sufficient cell comparisons; recombinant GCase uptake/trafficking assay |
FASEB journal |
Medium |
26018676
|
| 2015 |
LIMP-2 is a substrate of cathepsin-F: cathepsin-F mediates proteolytic cleavage of wild-type LIMP-2 in lysosomes in vitro and in vivo. Disease-causing cathepsin-F mutants (associated with type-B Kufs disease) fail to cleave LIMP-2. |
Heterologous expression of cathepsin-F variants, in vitro cleavage assay, purified lysosomes from mouse tissue, immunoblotting |
Biochemical and biophysical research communications |
Medium |
25576872
|
| 2010 |
LIMP-2 controls late phagosomal trafficking and the innate immune response to Listeria monocytogenes in macrophages: LIMP-2-deficient mice show impaired phago-lysosome transformation, low listericidal activity, reduced acute-phase pro-inflammatory cytokines, and 25-fold increased susceptibility to Listeria. LIMP-2 transfection in CHO cells confirms its role in late endosomal/lysosomal fusion and activation of Rab5a. |
LIMP-2-deficient mouse macrophage functional assays, cytokine/chemokine measurement, phagolysosome biogenesis assay, CHO cell reconstitution with LIMP-2 transfection, Rab5a activation assay |
The Journal of biological chemistry |
Medium |
21123180
|
| 2014 |
LIMP-2 expression is critical for lysosomal GCase activity and α-synuclein clearance: LIMP-2-deficient mouse brains show reduced GCase activity, lipid storage, disturbed autophagy/lysosomal function, and α-synuclein accumulation causing dopaminergic neuron toxicity. Heterologous expression of LIMP-2 accelerates clearance of overexpressed α-synuclein by increasing lysosomal GCase activity. |
LIMP-2-deficient mouse brain analysis, GCase activity assay, autophagy markers, α-synuclein immunofluorescence/immunoblot, LIMP-2 overexpression rescue experiment |
Proceedings of the National Academy of Sciences of the United States of America |
High |
25316793
|
| 2011 |
LIMP-2 deficiency causes a defect in proteolysis of reabsorbed proteins in renal proximal tubule cells: megalin/cubilin-dependent endocytosis is unaffected, but cathepsin B fails to co-localize with endosomal contents in Limp-2−/− mice, indicating that LIMP-2 is required for fusion of endosomes with lysosomes in the proximal tubule. |
Limp-2 knockout mice, in vivo fluorescent albumin uptake/tracking, cathepsin B co-localization by immunofluorescence, megalin/cubilin expression analysis |
American journal of physiology. Renal physiology |
Medium |
21429972
|
| 2024 |
GSH (glutathione) directly binds to SCARB2, interfering with the interaction between its N and C termini. This recruits mTORC1 to lysosomes through ARF1, leading to activation of mTOR signaling and promoting breast cancer progression. |
TME metabolomics, GCLC adipocyte-specific knockout mouse model, direct GSH-SCARB2 binding assay, mTORC1 lysosomal recruitment assay, ARF1 interaction studies |
Cell metabolism |
Medium |
39442522
|
| 2023 |
SCARB2 promotes MYC acetylation by interfering with HDAC3-mediated deacetylation of MYC at lysine 148, thereby enhancing MYC transcriptional activity and hepatocellular carcinoma cancer stem cell properties. Knockout of Scarb2 in hepatocytes attenuates HCC initiation in MYC-driven and DEN-induced mouse models. |
CRISPR/Cas9 knockout library screen in HCC tumorspheres, Scarb2 hepatocyte-specific knockout HCC mouse models, co-immunoprecipitation of SCARB2/MYC/HDAC3, MYC acetylation assay at K148 |
Nature communications |
Medium |
37739936
|
| 2024 |
Cryo-EM structure of GCase in complex with LIMP-2 reveals that helix 5 and helix 7 of LIMP-2's ectodomain interact with a binding pocket on GCase via a mostly hydrophobic interface supported by one essential salt bridge. LIMP-2 overexpression increases lysosomal abundance and enzymatic activity of GCase, and acts as an allosteric activator; a peptide derived from the LIMP-2 single helix enhances lysosomal GCase activity in patient-derived fibroblasts. |
Cryo-electron microscopy with engineered LIMP-2 shuttle and pro-macrobodies, LIMP-2 overexpression in HEK293T cells, GCase activity assays in fibroblasts, co-purification of GCase-LIMP-2 complex |
Nature communications |
High |
40159502
|
| 2024 |
LIMP-2 is present at ER-lysosome membrane contact sites through interaction with the endosomal protein STARD3 and ER-resident VAPB; STARD3 is required for the LIMP-2/VAPB interaction. This places LIMP-2 at organelle contact sites that may facilitate cholesterol transport from lysosomal to ER membrane. |
Proximity-based interaction screen (BioID), co-immunoprecipitation, immunofluorescence co-localization |
Journal of cell science |
Medium |
39370902
|
| 2024 |
Tetrandrine directly binds the LIMP-2 ectodomain (identified by clickable photoaffinity probe), inhibiting lysosomal cholesterol and sphingosine transport. LIMP-2 depletion or tetrandrine treatment inhibits NAADP-dependent calcium release via two-pore channels (TPCs); this is reversed by removing lysosomal cholesterol and sphingosine. Sphingosine triggers TPC-mediated lysosomal calcium release and restores this signaling in LIMP-2-deficient cells. |
Clickable photoaffinity probe for target identification, LIMP-2 knockdown, lysosomal calcium assays, sphingosine supplementation rescue, TPC functional assays |
Nature communications |
Medium |
40628771
|
| 2024 |
SCARB2 deficiency in mice leads to gut dysbiosis and altered bile acid pool, causing hyperactivation of intestinal FXR, which impairs epithelium renewal and dietary lipid (including vitamin E) absorption. FXR inhibition or vitamin E supplementation ameliorates neuromotor impairment and neuropathy in Scarb2 knockout mice. |
Scarb2 knockout mouse model, gut microbiome analysis, bile acid profiling, FXR activity assay, vitamin E level measurement in patients, FXR inhibitor and vitamin E supplementation rescue experiments |
Protein & cell |
Medium |
38635907
|
| 2024 |
EV-A71 does not encounter its uncoating receptor SCARB2 at the cell surface; SCARB2 is absent from the surface of RD and other susceptible cell lines and is concentrated in lysosomes/late endosomes. SCARB2 is dispensable for virus attachment but essential for infection, indicating that the critical SCARB2-EV-A71 interaction occurs intracellularly (in lysosomes) to trigger uncoating rather than at the plasma membrane. |
SCARB2 and PSGL-1 knockout cell lines, cell surface expression analysis, virus attachment assays, infection assays |
PLoS pathogens |
Medium |
38359079
|
| 2003 |
LIMP-2/LGP85-deficient mice exhibit peripheral demyelinating neuropathy associated with massive loss of peripheral myelin proteins and increased lysosomal protein activity, indicating a role for the lysosomal compartment in peripheral myelination; mice also show lysosome accumulation in ureteric epithelium with disturbed uroplakin surface expression, causing ureteropelvic junction obstruction. |
LIMP-2 knockout mouse phenotypic analysis: neuropathology, myelin protein immunoblotting, lysosomal enzyme activity assays, histology, immunofluorescence |
Human molecular genetics |
Medium |
12620969
|
| 2022 |
SR-B2/LIMP-2 is present at the RPE cell surface (not exclusively intracellular), associates with lipid rafts, and participates in the control of photoreceptor outer segment (POS) phagocytosis speed in retinal pigment epithelial cells; siRNA inhibition of SR-B2/LIMP-2 alters POS internalization dynamics, similar to CD36. |
siRNA knockdown, immunoblotting, immunohistochemistry, lipid raft flotation gradients, phagocytosis assays in RPE cell lines and tissue |
International journal of molecular sciences |
Low |
35408805
|
| 2019 |
In a human iPSC-derived cardiomyocyte model of Fabry disease, LIMP-2 accumulates relative to controls; overexpression of LIMP-2 directly induces secretion of cathepsin F and HSPA2/HSP70-2 and causes massive vacuole accumulation, suggesting LIMP-2 accumulation is causally linked to these downstream pathological events. |
iPSC-derived cardiomyocytes from Fabry disease patients, quantitative proteomics, LIMP-2 overexpression with cathepsin F/HSP70-2 secretion readout, genetic correction reversal |
Stem cell reports |
Medium |
31378672
|
| 2022 |
SCARB2 deficiency disrupts mTORC1-dependent mitochondrial oxidative phosphorylation (OXPHOS) in adipocytes: Scarb2 deficiency decreases the mTORC1/4E-BP1 pathway, leading to impaired mitochondrial respiration and enhanced glycolysis, resulting in reduced lipid storage in white adipose tissue. |
Adiponectin-Cre; Scarb2 conditional knockout mice, mTORC1/4E-BP1 pathway analysis by immunoblot, mitochondrial respiration (Seahorse), glycolysis assay |
International journal of molecular sciences |
Medium |
35955761
|