| 2000 |
NEU3 encodes a 428-residue human plasma membrane-associated sialidase with a putative transmembrane helix, YRIP motif, and three Asp boxes; expressed in COS7 cells it shows high ganglioside-hydrolyzing sialidase activity (pH optimum 3.8) and localizes to the plasma membrane by immunofluorescence. |
cDNA cloning, transient transfection in COS7 cells, sialidase activity assay, immunofluorescence |
The Biochemical journal |
High |
10861246
|
| 2002 |
NEU3 is enriched in caveolae microdomains and associates directly with caveolin-1 via a caveolin-binding motif; a single amino acid mutation in this motif reduces microdomain recruitment and sialidase activity; caveolin-1 co-elutes with His-tagged NEU3 on affinity chromatography and co-immunoprecipitates with anti-caveolin-1 antibody; cholesterol depletion displaces NEU3 from microdomains and decreases activity, while increased caveolin-1 expression activates NEU3. |
Sucrose density gradient fractionation, affinity column chromatography, co-immunoprecipitation, site-directed mutagenesis, beta-cyclodextrin cholesterol depletion, immunoblotting |
The Journal of biological chemistry |
High |
12011038
|
| 2002 |
NEU3 overexpression in human colon cancer cells inhibits apoptosis, accompanied by increased Bcl-2 and decreased caspase expression; NEU3 is downregulated during sodium butyrate-induced differentiation/apoptosis; lactosylceramide, a NEU3 catalytic product, reduces apoptotic cells, suggesting NEU3 promotes survival through ganglioside modulation. |
Gene transfection into colon cancer cells, flow cytometry for apoptosis, western blot for Bcl-2/caspase, TLC for gangliosides, sodium butyrate treatment |
Proceedings of the National Academy of Sciences of the United States of America |
High |
12149448
|
| 2006 |
NEU3 overexpression in colon cancer DLD-1 cells increases adhesion to laminins and cell proliferation, but decreases adhesion to fibronectin; on laminin-5, NEU3 stimulates phosphorylation of FAK and ERK, co-immunoprecipitates with integrin β4, recruits Shc and Grb-2 to integrin β4, and promotes phosphorylation of integrin β1 and ILK; GM3 depletion by NEU3 correlates with these bimodal adhesion effects. |
Transfection, co-immunoprecipitation with anti-integrin β4 antibody, western blot for phospho-FAK/ERK/integrin β1/ILK, cell adhesion assay, TLC for gangliosides |
The Biochemical journal |
High |
16241905
|
| 2006 |
EGF stimulation redistributes NEU3 to membrane ruffles where it co-localizes and co-precipitates with activated Rac-1; NEU3 overexpression enhances Rac-1 activation and cell migration, while NEU3 siRNA silencing inhibits Rac-1 activation. |
Immunofluorescence, GST-PAK-1 pulldown for activated Rac-1, siRNA knockdown, migration assay |
Biochemical and biophysical research communications |
High |
16765317
|
| 2007 |
Hepatic overexpression of NEU3 via adenoviral vectors improves insulin sensitivity and glucose tolerance in mice through modification of ganglioside composition (increased GM1, markedly reduced GM3 in liver); increases tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1) but not insulin receptor or IRS-2; enhances PPARγ and fetuin expression. |
Adenoviral gene delivery in mice, TLC for gangliosides, insulin tolerance test, glucose tolerance test, western blot for phospho-IRS-1 |
Metabolism: clinical and experimental |
High |
17292733
|
| 2007 |
NEU3 subcellular localization was determined: NEU3 is a peripheral membrane protein associated with the outer leaflet of the plasma membrane (demonstrated by surface biotinylation and carbonate extraction), is also present in a subset of endosomal compartments, is internalized from the plasma membrane and sorted to recycling endosomes, and is hydrophilic as shown by Triton X-114 phase separation. |
Selective cell-surface protein biotinylation, carbonate extraction, Triton X-114 phase separation, immunofluorescence, subcellular fractionation |
The Biochemical journal |
High |
17708748
|
| 2008 |
NEU3 silencing in C2C12 myoblasts increases GM3 ganglioside above a critical threshold, causing EGFR inhibition and eventual EGFR down-regulation, which blocks myoblast differentiation and increases apoptosis susceptibility; NEU3 strictly modulates the GM3 content of adjacent cells and is required for myogenic differentiation. |
siRNA-mediated NEU3 knockdown, TLC for gangliosides, western blot for EGFR, differentiation assay, apoptosis assay |
The Journal of biological chemistry |
High |
18945680
|
| 2008 |
NEU3 silencing in K562 chronic myeloid leukemia cells markedly increases GM3 and other gangliosides, reduces cell growth, shifts cell cycle regulators (decreases cyclin D2 and Myc, increases p21), promotes apoptosis susceptibility, and triggers megakaryocytic differentiation; downstream signaling through PLC-β2, PKC, RAF, ERK1/2, RSK90, and JNK is activated upon NEU3 silencing. |
siRNA knockdown, flow cytometry, TLC for gangliosides, RT-PCR, western blot for signaling molecules |
Cell death and differentiation |
High |
18820643
|
| 2009 |
NEU3 transgenic mice show significantly increased azoxymethane-induced colonic aberrant crypt foci formation associated with enhanced phosphorylation of EGFR, Akt and ERK, up-regulation of Bcl-xL, reduced apoptosis (assessed by cleaved caspase-3), and decreased GM3 with increased lactosylceramide in colonic mucosa. |
Transgenic mouse model, azoxymethane treatment, immunohistochemistry for cleaved caspase-3, western blot for phospho-EGFR/Akt/ERK/Bcl-xL, TLC for gangliosides |
Cancer science |
High |
19215228
|
| 2010 |
Site-directed mutagenesis of recombinant human NEU3 identified key catalytic residues: general acid-base D50 and nucleophilic Y370-E225 pair; NMR spectroscopy confirmed NEU3 acts as a retaining exo-sialidase; residues A160, M87, I105 interact with the N5-acetyl group and E113, Y179, Y181 with the C7-C9 glycerol side-chain of sialic acid; N- or C-terminal truncations >10 residues abolish activity. |
Site-directed mutagenesis, in vitro enzymatic assays, NMR spectroscopy, homology modeling |
Glycobiology |
High |
20511247
|
| 2011 |
NEU3 overexpression in prostate cancer LNCaP cells induces expression of EGR-1, androgen receptor (AR) and PSA both with and without androgen; this induction is abrogated by PI3K and MAP kinase inhibitors and confirmed by increased AKT and ERK1/2 phosphorylation; NEU3 silencing reduces growth of androgen-independent PC-3 cells in culture and in nude mice xenografts. |
siRNA knockdown, forced overexpression, western blot for phospho-AKT/ERK, kinase inhibitors (PI3K, MAPK), xenograft tumor assay |
Cell death and differentiation |
High |
21681193
|
| 2011 |
NEU3 substrate recognition requires a hydrophobic aglycone; enzymatic activity is directly dependent on aglycone hydrophobicity but not on features of the ceramide headgroup; azide modifications at C9 or N5-Ac of Neu5Ac are tolerated, but large aryl groups are accepted only at C9 and not at N5-Ac; a two-site recognition model (Neu5Ac binding + hydrophobic aglycone interaction) is proposed. |
Electrospray ionization mass spectrometry-based substrate cleavage assay with synthetic trisaccharide substrates |
Biochemistry |
High |
21675735
|
| 2012 |
NEU3 silencing in renal carcinoma cells increases membrane ganglioside content (especially GD1a), up-regulates RAB25 (directing integrins to lysosomes), down-regulates CLIC3 (which recycles integrins to plasma membrane), enhances caveolar endocytosis of β1 integrin, blocks β1 integrin recycling to the plasma membrane, and consequently inhibits EGFR and FAK/AKT signaling. |
siRNA knockdown, western blot for RAB25/CLIC3/EGFR/FAK/AKT, flow cytometry for surface integrin, TLC for gangliosides, invasion/adhesion assays |
The Journal of biological chemistry |
High |
23139422
|
| 2012 |
NEU3 silencing in skeletal muscle C2C12 cells under hypoxia increases apoptosis; NEU3 up-regulation under hypoxia stimulates EGFR signaling which activates HIF-1α, increasing cell survival and proliferation; stable NEU3 overexpression enhances hypoxia resistance while stable silencing increases apoptosis susceptibility. |
Stable overexpression and siRNA silencing, western blot for HIF-1α/phospho-EGFR, hypoxia treatment, apoptosis assay |
The Journal of biological chemistry |
High |
23209287
|
| 2014 |
In adult rat PNS sensory axons, axotomy activates Neu3 sialidase via calcium influx that activates P38MAPK, which then activates Neu3; this converts GD1a and GT1b to GM1, which is essential for axon regeneration; externally applied sialidase rescues regeneration in CNS axons where Neu3 is not activated by injury; Neu3 activation further stimulates the ERK pathway. |
In vitro and in vivo axotomy models (rat DRG and retinal axons), calcium imaging, pharmacological inhibitors of P38MAPK and Neu3 sialidase, TLC for gangliosides, exogenous sialidase application |
The Journal of neuroscience |
High |
24523539
|
| 2014 |
STD NMR with catalytically inactive NEU3(Y370F) confirmed close contacts between the enzyme and both the hydrophobic aglycone and the Neu5Ac residue of GM3-analog substrates; facial recognition of galactose and glucose residues was identified; molecular dynamics simulations corroborated the homology model predictions. |
STD NMR spectroscopy with inactive mutant NEU3(Y370F)-MBP fusion protein, molecular dynamics simulations |
Glycobiology |
High |
25294388
|
| 2015 |
Phosphatidic acid (PA) produced by phospholipase D1 (PLD1) directly activates NEU3 sialidase activity 4–5-fold in vitro and promotes its translocation to the cell surface; NEU3 interacts selectively with PA (phospholipid array, liposome co-precipitation, ELISA); PA- and calmodulin-binding sites were mapped to the N-terminal region; EGF induces PLD1 activation concomitant with NEU3 translocation; NEU3-PA interaction promotes cell migration through Ras signaling. |
In vitro sialidase activity assay with phospholipids, phospholipid array, liposome co-precipitation, ELISA, confocal microscopy, site-directed mutagenesis of N-terminal fragments, migration assay |
FASEB journal |
High |
25678627
|
| 2015 |
NEU3 (active form but not inactive mutant) co-immunoprecipitates with EGFR and Src kinase; NEU3 activates Src kinase activity; EGFR/Src pathway activation by NEU3 promotes oncogenic transformation (clonogenicity on soft agar, in vivo tumor growth); Src inhibitor PP2 completely suppresses NEU3-mediated clonogenicity; activity-null mutants fail to activate Src or EGFR, indicating ganglioside modulation is required. |
Co-immunoprecipitation, Src kinase activity assay, soft agar clonogenicity, nude mouse xenograft, EGFR and Src inhibitors, activity-null mutant comparison |
PloS one |
High |
25803810
|
| 2015 |
Active NEU3 (not inactive mutant) enhances EGFR activation (hyperphosphorylation) without affecting EGFR mRNA or protein expression; EGFR immunoprecipitated from NEU3-overexpressing cells is desialylated as shown by mass spectrometry and western blot; NEU3 thus activates EGFR both indirectly (via GM3 reduction) and directly (via EGFR desialylation). |
Transfection of wild-type vs. inactive mutant NEU3, co-immunoprecipitation, mass spectrometry, western blot for phospho-EGFR |
Glycobiology |
High |
25922362
|
| 2015 |
NEU3 silencing in colon cancer HT-29 and HCT116 cells decreases phosphorylation of LRP6 (Wnt co-receptor), reduces β-catenin activation, impairs LRP6 complex formation with GSK3β and Axin, and reduces clonogenicity and in vivo tumor growth; activity-null NEU3 mutant fails to activate Wnt signaling; NEU3 also regulates ERK and Akt phosphorylation via EGFR/Ras. |
siRNA knockdown, activity-null mutant, western blot for phospho-LRP6/β-catenin/GSK3β/ERK/Akt, immunoprecipitation for LRP6 complex, soft agar assay, xenograft in NOD-SCID mice |
International journal of cancer |
High |
25810027
|
| 2015 |
NEU3 expression reduces transferrin (Tf) internalization via clathrin-mediated endocytosis (CME); NEU3 decreases internalization of α2-macroglobulin and LDL (other CME ligands) but not cholera toxin β-subunit; NEU3 reduces Tf sorting to early and recycling endosomes and decreases Tf binding at cell surface; NEU3-expressing cells show altered subcellular distribution of clathrin adaptor AP-2 but not clathrin, PtdIns(4,5)P2, or caveolin-1. |
Ectopic NEU3 expression, fluorescence-based Tf internalization assay, confocal microscopy for AP-2/clathrin distribution, glycosphingolipid depletion experiments |
The Biochemical journal |
High |
26251452
|
| 2016 |
NEU3 protein interactors were identified by mass spectrometry in plasma membrane and endosomal compartments; NEU3 localizes dynamically between plasma membrane (high activity) and endosomes (low activity); under appropriate stimuli NEU3 shifts from endosomes to plasma membrane with increased activity; selected interactors were validated by cross-immunoprecipitation. |
Mass spectrometry proteomics, cross-immunoprecipitation, subcellular fractionation, enzyme activity assay in fractions |
The Journal of biological chemistry |
Medium |
26987901
|
| 2017 |
Human NEU3 is S-acylated (palmitoylated) on its cytosolic-exposed C-terminal domain; NEU3 behaves like an integral membrane protein (not released by conditions that extract peripheral proteins), with C-terminus exposed to cytosol and another portion exposed extracellularly; in silico analysis and homology modeling indicate no α-helical transmembrane segment, suggesting S-acylation contributes to membrane anchorage. |
S-acylation biochemical assay, carbonate extraction, topology probing by protease protection and selective biotinylation, homology modeling |
Scientific reports |
High |
28646141
|
| 2017 |
NEU3 overexpression in glioblastoma cells reduces invasion and migration by promoting focal adhesion assembly through reduced calpain-dependent proteolysis; NEU3 silencing elevates calpain activity and GM3 accumulation, and localizes calpain and GM3 to the cell lamellipodium; activity-null NEU3 fails to reduce invasion, indicating ganglioside hydrolysis is required. |
Transwell invasion/migration assay, western blot for calpain and focal adhesion proteins, immunofluorescence microscopy, siRNA knockdown and overexpression |
Biochimica et biophysica acta. General subjects |
High |
28760640
|
| 2017 |
NEU3 is associated with the outer leaflet of exosomes secreted by HeLa cells and retains enzymatic activity on the exosome surface; NEU3 localization on exosomes was confirmed by enzyme activity measurements, western blot, and dot blot. |
Inducible NEU3 expression in HeLa cells, exosome purification, enzyme activity assay on exosomes, western blot, dot blot |
Biochemistry |
Medium |
29039925
|
| 2017 |
Murine Neu3 is responsible for GM2 ganglioside catabolism in the mouse brain as a bypass of HEXA deficiency; Hexa-/-Neu3-/- double knockout mice accumulate GM2 ganglioside in brain, develop neurodegeneration, ataxia, and die at 1.5–4.5 months, recapitulating Tay-Sachs disease; Neu3 sialidase converts GM2 to GA2 allowing further processing by β-hexosaminidase B. |
Double knockout mouse generation, TLC, mass spectrometry for gangliosides, histology, immunohistochemistry, electron microscopy, behavioral testing |
Experimental neurology |
High |
28974375
|
| 2010 |
Sp1 and Sp3 transcription factors bind the NEU3 gene promoter and regulate its expression; the NEU3 gene has alternative promoters with two clusters of transcription start sites — one preferentially used in brain and another in other tissues; Sp1/Sp3 siRNA knockdown differentially modulates these promoters, increasing brain-type transcription while decreasing transcription from other TSSs. |
Oligo-capping for TSS mapping, luciferase reporter assay, electrophoretic mobility-shift assay (EMSA), chromatin immunoprecipitation (ChIP), siRNA knockdown of Sp1/Sp3 |
The Biochemical journal |
High |
20518744
|
| 2014 |
NEU3 sialidase activity in DRM (lipid raft) and non-DRM membrane compartments differentially modifies ganglioside composition in each compartment; newly synthesized NEU3 associates first with non-DRM, then redistributes to both DRM and non-DRM at steady state; NEU3 is degraded via the proteasomal pathway; NEU3 triggers Akt phosphorylation even without exogenous EGF. |
Inducible expression system, density gradient fractionation, TLC for gangliosides, proteasome inhibitor treatment, western blot for phospho-Akt |
PloS one |
Medium |
24925219
|
| 2021 |
Mammalian NEU3 neuraminidase is responsible for intestinal desialylation of alkaline phosphatase (IAP) during Salmonella-induced colitis; absence of NEU3 prevents IAP desialylation, prevents LPS-phosphate accumulation, and prevents inflammatory cytokine expression, thereby protecting against severe colitis development. |
Neu3 knockout mouse model, intestinal IAP activity assay, LPS-phosphate measurement, cytokine quantification, colitis model |
Proceedings of the National Academy of Sciences of the United States of America |
High |
34266954
|
| 2022 |
Recombinant active NEU3 (but not catalytically inactive NEU3) is sufficient to induce pulmonary fibrosis and inflammation when administered by oropharyngeal aspiration in mice; NEU3 knockout mice show strongly attenuated bleomycin-induced fibrosis, indicating NEU3 is both necessary and sufficient for pulmonary fibrosis through its enzymatic activity. |
Recombinant NEU3 and inactive mutant aspiration, Neu3 knockout mice, bleomycin fibrosis model, histology, BAL fluid analysis |
Respiratory research |
High |
35999554
|
| 2022 |
NEU3 (but not NEU1, NEU2, or NEU4) primes human neutrophils: extracellular NEU3 induces amoeboid morphology, redistributes primed neutrophil markers CD11b, CD18, and CD66a to the cell cortex, decreases CD43 and CD62-L at cortex, and increases F-actin content; NEU3 effect depends on its enzymatic activity (blocked by NEU3 inhibitor 2-acetylpyridine) and is associated with cell surface desialylation. |
Recombinant NEU3 incubation with human neutrophils, flow cytometry, confocal microscopy for F-actin and surface markers, pharmacological inhibition with 2-acetylpyridine |
Journal of leukocyte biology |
High |
35899930
|
| 2022 |
NEU3 activity influences CD22 cluster size on B cells; NEU3 activity increases lateral mobility of CD22 (measured by single-particle tracking), in contrast to exogenous bacterial neuraminidase; native NEU1 and NEU3 activities influence cellular Ca2+ levels in B cells, supporting a role in B cell activation regulation. |
Confocal microscopy for CD22 clustering, single-particle tracking, Ca2+ measurement, pharmacological sialidase inhibition |
Biophysical reports |
Medium |
36425332
|
| 2023 |
NEU3 acts on GM1 ganglioside to produce GA1 glycolipid in the mouse brain, providing a bypass catabolism route in the absence of β-galactosidase (Glb1); Glb1/Neu3 double KO mice accumulate more GM1 and less GA1 compared to Glb1 single KO, develop more severe neurodegeneration and ataxia with shorter lifespan; mouse NEU3 converts GM1 to GA1 more efficiently than human NEU3. |
Double knockout mouse generation, lipidomics (TLC, mass spectrometry), behavioral testing, neuropathology, interspecies enzyme activity comparison |
Journal of lipid research |
High |
37871851
|
| 2020 |
Induced NEU3 overexpression in primary human cardiac fibroblasts reduces ganglioside GM3 content and significantly reduces TGF-β signaling pathway activation, ultimately decreasing collagen I deposition; NEU3 acts as an inhibitor of cardiac fibroblast activation through GM3 modulation. |
Forced NEU3 overexpression in primary human cardiac fibroblasts, TLC for gangliosides, western blot for TGF-β pathway components, collagen I measurement |
The Biochemical journal |
Medium |
32869836
|
| 2025 |
TGF-β1 rapidly increases NEU3 protein levels (within 5 minutes) in human lung fibroblasts through increased translation (not transcription); the RNA helicase DDX3 mediates NEU3 translation; TGF-β1 induces DDX3 dephosphorylation within 2 minutes; DDX3 inhibitors block rapid NEU3 upregulation; NEU3 activates latent TGF-β1 by cleaving sialic acid from the LAP peptide, creating a positive feedback loop; NEU3 inhibitors block this feedback loop. |
Time-course protein expression, transcription/translation inhibitor experiments, DDX3 inhibitors, phosphorylation analysis, TGF-β1 activation assay, NEU3 inhibitors |
bioRxivpreprint |
Medium |
bio_10.1101_2025.10.16.682941
|