Affinage

NGLY1

Peptide-N(4)-(N-acetyl-beta-glucosaminyl)asparagine amidase · UniProt Q96IV0

Length
654 aa
Mass
74.4 kDa
Annotated
2026-06-10
97 papers in source corpus 26 papers cited in narrative 27 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/8 claims corpus-supported (88%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NGLY1 is a soluble cytosolic/nuclear peptide:N-glycanase that drives the cytoplasmic deglycosylation arm of ER-associated degradation (ERAD), removing N-linked glycans from misfolded glycoproteins retrotranslocated from the ER and coupling their turnover to the proteasome (PMID:10831608, PMID:16401726). Rather than simply excising sugars, the enzyme performs 'sequence editing', deamidating the glycosylated asparagine to aspartate and thereby introducing a permanent negative charge into the substrate peptide (PMID:38581946). It is escorted to the 26S proteasome through a direct physical interaction with the ubiquitin-binding protein RAD23, which links deglycosylated substrates to degradation (PMID:11259433, PMID:16401726). A central physiological output of this activity is activation of the transcription factor NFE2L1/NRF1: NGLY1-mediated processing of retrotranslocated NRF1 is strictly required for its nuclear entry and induction of the proteasome 'bounce-back' response (PMID:29202016), and this axis extends to mitochondrial homeostasis and mitophagy, ferroptosis resistance via GPX4, and in vivo hepatic NRF1 function (PMID:30135079, PMID:35271393, PMID:31733337). NGLY1 also acts through NRF1-independent routes, maintaining AMPKα levels and energy metabolism across fly, mouse, and patient cells (PMID:33315951). In NGLY1 deficiency the alternative deglycosidase ENGase generates aggregation-prone N-GlcNAc proteins that are the primary pathological drivers, since deleting ENGase restores ERAD and partially rescues lethality (PMID:25605922, PMID:28426790). Loss of NGLY1 produces neuronal protein aggregation, mitochondrial and synaptic defects in human iPSC-derived cortical neurons (PMID:38039131), and STING-dependent neurodegeneration in mice that is rescued by Sting1 ablation or pharmacological STING inhibition (PMID:40644312, PMID:30135079). Beyond ERAD, NGLY1 has enzyme-independent scaffolding roles in cell polarity and neuronal branching in invertebrate orthologs (PMID:19940117, PMID:20130186).

Mechanistic history

Synthesis pass · year-by-year structured walk · 25 steps
  1. 2000 High

    Established the core biochemical identity: NGLY1's yeast ortholog is a soluble cytosolic/nuclear enzyme with intrinsic PNGase activity whose loss impairs degradation of a misfolded glycoprotein, defining a deglycosylation-to-degradation function.

    Evidence Genetic PNGase-deficiency screen, recombinant E. coli expression with enzymatic assay, and subcellular fractionation in yeast

    PMID:10831608

    Open questions at the time
    • Did not identify the proteasome-coupling partner
    • Mammalian substrate repertoire undefined
  2. 2001 High

    Answered how deglycosylation is physically linked to degradation by identifying a Png1p-Rad23p complex distinct from the DNA-repair Rad4p-Rad23p complex, casting Rad23 as the escort to the 26S proteasome.

    Evidence Yeast two-hybrid screen with reciprocal co-immunoprecipitation

    PMID:11259433

    Open questions at the time
    • Did not show requirement of the complex for substrate turnover
    • Cofactor determinants of substrate specificity unknown
  3. 2006 High

    Demonstrated functional necessity of the complex: Png1-Rad23 is required for efficient degradation of a glycosylated ERAD substrate, establishing the complex as the operative unit coupling deglycosylation to proteolysis.

    Evidence Genetic deletion of PNG1/RAD23 with glycoprotein (ricin A chain) turnover assays in yeast

    PMID:16401726

    Open questions at the time
    • Limited to a model substrate
    • No mammalian validation at this stage
  4. 2009 Medium

    Revealed that NGLY1 has functions independent of its catalytic activity, since a catalytically dead ortholog is still essential for cell polarity, decoupling scaffolding roles from deglycosylation.

    Evidence Gene deletion and catalytic-mutant morphological analysis in Neurospora crassa

    PMID:19940117

    Open questions at the time
    • Single organism, single lab
    • Molecular basis of the non-catalytic role unresolved
    • Relevance to mammalian NGLY1 untested
  5. 2010 High

    Placed NGLY1 in neuronal morphogenesis, showing png-1 acts with rad-23 in the same pathway to restrict axon branching, linking the deglycosylation/degradation axis to nervous-system development.

    Evidence C. elegans loss-of-function screen, tissue-specific rescue, and genetic epistasis with rad-23

    PMID:20130186

    Open questions at the time
    • Substrate driving branching unknown
    • Catalytic vs scaffolding dependence not dissected
  6. 2014 Medium

    Showed that cytosolic versus ER-membrane-associated pools of Png1 have distinct, even opposing, functional consequences on the same substrate class, refining where and how deglycosylation acts.

    Evidence Yeast PNG1 deletion with ricin A chain trafficking, depurination, and toxicity assays

    PMID:25436896

    Open questions at the time
    • Substrate-specific generalization unclear
    • Mechanism of pool partitioning undefined
  7. 2015 High

    Identified the pathological mechanism of NGLY1 loss: in its absence ENGase generates aggregation-prone N-GlcNAc proteins, and removing ENGase restores ERAD, defining a druggable disease driver.

    Evidence Ngly1-/- and Ngly1-/-/ENGase-/- MEFs with degradation and aggregation assays

    PMID:25605922

    Open questions at the time
    • Organismal relevance not yet established at this stage
    • Identity of key aggregating substrates undefined
  8. 2017 High

    Defined a major physiological output by showing NGLY1 deglycosylation of retrotranslocated NRF1 is required for its nuclear translocation and the proteasome bounce-back response, linking NGLY1 to proteostatic transcriptional control.

    Evidence Genetic KO, siRNA, a small-molecule NGLY1 inhibitor, and NRF1 localization/transcription reporter assays in human cells

    PMID:29202016

    Open questions at the time
    • Full NRF1 target program not delineated
    • Tissue-level consequences untested at this stage
  9. 2017 High

    Confirmed the ENGase epistasis at the organismal level: ENGase deletion partially rescues Ngly1-/- mouse lethality, validating ENGase-generated N-GlcNAc aggregates as primary disease drivers in vivo.

    Evidence Ngly1-/- x ENGase-/- double-knockout mouse survival analysis

    PMID:28426790

    Open questions at the time
    • Rescue only partial, implying additional pathology
    • Neurological phenotypes not fully resolved
  10. 2018 High

    Extended NGLY1 function to organelle quality control and innate immunity, showing NGLY1-NRF1 maintains mitophagy and that its loss chronically activates cGAS-STING/MDA5-MAVS, raising an interferon signature rescuable by NRF2 activation.

    Evidence Human and mouse NGLY1-deficient cells with mitophagy, mitochondrial, and nucleic-acid-sensing pathway assays plus NRF2-activator rescue

    PMID:30135079

    Open questions at the time
    • In vivo contribution of STING not yet tested at this stage
    • Trigger linking mitochondrial fragmentation to sensing undefined
  11. 2018 Medium

    Linked NGLY1 loss to neuroendocrine signaling and confirmed an NRF1 dysfunction signature without ER stress in flies, with ecdysone rescue of developmental delay.

    Evidence Drosophila Pngl loss-of-function model with RNAseq, tissue-specific rescue, and 20-hydroxyecdysone supplementation

    PMID:29346549 PMID:29735526

    Open questions at the time
    • Mechanism connecting NGLY1 to ecdysteroid production unclear
    • Mammalian relevance of neuroendocrine axis untested
  12. 2019 Medium

    Established tissue-level in vivo requirement for NGLY1 in NRF1 processing, with hepatocyte-specific loss causing impaired NRF1 nuclear localization and lipid/nuclear-morphology defects under dietary stress.

    Evidence Liver-specific conditional Ngly1 knockout mice with NRF1 processing and liver phenotype analysis

    PMID:31733337

    Open questions at the time
    • Single tissue, single lab
    • Contribution relative to ENGase aggregates unquantified
  13. 2020 High

    Defined an NRF1-independent NGLY1 pathway: NGLY1 loss reduces AMPKα and disrupts energy metabolism across fly, mouse, and patient cells, with epistasis excluding NFE2L1 as the cause.

    Evidence Drosophila tissue-specific genetics, Ngly1-/- MEFs, patient fibroblasts, AMPKα western blot, pharmacological AMPK activation, and NFE2L1 epistasis

    PMID:33315951

    Open questions at the time
    • Molecular basis of AMPKα reduction undefined
    • Whether AMPKα is a direct substrate unknown
  14. 2020 Medium

    Identified candidate functional substrate/modifier NKCC1, whose molecular weight and function are altered in NGLY1-deficient cells, expanding the substrate landscape beyond NRF1.

    Evidence Drosophila genetic diversity panel association plus NGLY1-/- mouse cell NKCC1 functional assays

    PMID:33315011

    Open questions at the time
    • Direct deglycosylation of NKCC1 not demonstrated
    • Mechanism of functional dependence unresolved
  15. 2022 Medium

    Connected NGLY1 to ferroptosis resistance, showing NGLY1-dependent NFE2L1 processing maintains GPX4 expression independently of NRF2, defining a distinct cell-survival axis.

    Evidence NGLY1/NFE2L1 KO cell lines with ferroptosis assays, GPX4 analysis, and NFE2L1/NFE2L2 epistasis

    PMID:35271393

    Open questions at the time
    • In vivo relevance untested
    • Single lab
  16. 2022 Medium

    Defined the chemistry of NRF1/SKN-1A activation as 'sequence editing' — Asn-to-Asp conversion strictly required for transcriptional activation — clarifying that NGLY1 alters substrate chemistry rather than merely stripping glycans.

    Evidence C. elegans genetics with transcriptomics and selective SKN-1A/SKN-1C mutant alleles (preprint)

    Open questions at the time
    • Preprint, not peer-reviewed
    • Generalization of sequence-editing requirement beyond SKN-1A undefined
  17. 2022 Medium

    Showed autophagy is a cytoprotective adaptation to NGLY1 loss, with autophagosome induction independent of ER stress, ROS, or Ca2+, and ATG13-dependence for survival.

    Evidence Pharmacological and siRNA NGLY1 inhibition with GFP-LC3 assays, autophagosome IP-MS, and ATG13-KO MEFs

    PMID:34995415

    Open questions at the time
    • Trigger of autophagy induction undefined
    • Pharmacological inhibitor specificity caveats
  18. 2023 High

    Established cell-type-specific neuronal vulnerability, with NGLY1-deficient human cortical neurons (not astrocytes) showing protein aggregation, mitochondrial, and synaptic defects rescued by NGLY1 reintroduction.

    Evidence Patient iPSC-derived cortical neurons with transcriptomics, proteomics, aggregate LCM-MS, and NGLY1 rescue

    PMID:38039131

    Open questions at the time
    • Causal substrate of aggregation unidentified
    • Why neurons are selectively vulnerable unresolved
  19. 2023 High

    Dissected organ-level NGLY1 loss in the gut, showing intestinal barrier defects and JNK/Foxo overactivation drive lethality through non-microbiotal immune/metabolic mechanisms.

    Evidence Tissue-specific Drosophila Pngl knockouts with germ-free rearing, dietary rescue, and JNK/Foxo pathway analysis

    PMID:37704604

    Open questions at the time
    • Mammalian relevance of gut phenotype untested
    • Direct NGLY1 substrates in this axis unknown
  20. 2024 Medium

    Identified an NRF1-independent metabolic modifier axis, showing nucleotide-metabolism gene mutations and nucleotide restriction suppress proteasome dysfunction in NGLY1-deficient worms.

    Evidence C. elegans suppressor screen, epistasis, and dietary nucleotide manipulation

    PMID:38991033

    Open questions at the time
    • Mechanism linking nucleotide availability to proteasome function unclear
    • Single organism
  21. 2024 Medium

    Provided practical and mechanistic confirmation of sequence editing, developing an ELISA neo-epitope assay detecting endogenous Asn-to-Asp conversion in patient iPSCs and demonstrating dynamic age-dependent NGLY1 activity in brain.

    Evidence ELISA neo-epitope activity assay in iPSC extracts and FRET-based probe assay in aging rat brain

    PMID:38417795 PMID:38581946

    Open questions at the time
    • Physiological drivers of age-dependent activity changes unknown
    • Substrate scope of endogenous editing unquantified
  22. 2024 Medium

    Expanded NGLY1's regulatory targets to immune checkpoint control, showing NGLY1 deglycosylates and destabilizes cancer-cell-intrinsic PD-1 upon doxorubicin to sensitize tumor cells.

    Evidence Co-IP, protein half-life assay, NGLY1 siRNA, and viability assays under doxorubicin

    PMID:38782868

    Open questions at the time
    • Single Co-IP context
    • In vivo tumor relevance untested
  23. 2024 High

    Provided structural context for the downstream fate of NGLY1-processed glycoproteins, resolving how the SCF-FBS2 ligase recognizes residual N-glycan remnants.

    Evidence X-ray crystallography of FBS2-SKP1-Man3GlcNAc2 with NMR and docking

    PMID:39171510

    Open questions at the time
    • Direct functional coupling to NGLY1 not demonstrated biochemically
    • Druggability not validated cellularly
  24. 2025 High

    Established the in vivo driver of NGLY1-deficiency neurodegeneration, showing STING ablation or inhibition rescues Purkinje cell loss and motor disease without inflammatory activation and via STING-dependent suppression of glial cholesterol biosynthesis.

    Evidence Postnatal inducible Ngly1 KO mice, Sting1 double-KO rescue, snRNA-seq, and pharmacological STING inhibition (VS-X4)

    PMID:40644312

    Open questions at the time
    • Upstream trigger activating STING in neurons undefined
    • Link between cholesterol suppression and degeneration mechanistic detail incomplete
  25. 2025 Medium

    Implicated NGLY1 in general ERAD capacity beyond specific-substrate deglycosylation, with SEL1L cytoplasmic-tail variants enhancing ERAD in an NGLY1-dependent manner and modifying lethality.

    Evidence CRISPR-engineered Drosophila SEL1L variants with ERAD, ER-stress, and proteasome-inhibition assays

    PMID:40773511

    Open questions at the time
    • Direct biochemical NGLY1-SEL1L interaction not shown
    • Mammalian validation absent

Open questions

Synthesis pass · forward-looking unresolved questions
  • How NGLY1's distinct catalytic modes (endo- vs exo-glycanase, sequence editing) and its enzyme-independent scaffolding functions are coordinated across substrates and tissues, and which substrates drive neuron-specific pathology, remains unresolved.
  • Exo-glycanase active site not validated by mutagenesis or structure
  • Comprehensive endogenous substrate catalog missing
  • Determinants of cell-type-specific vulnerability unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 5 GO:0016787 hydrolase activity 3 GO:0060089 molecular transducer activity 2 GO:0060090 molecular adaptor activity 2
Localization
GO:0005829 cytosol 2 GO:0005634 nucleus 1
Pathway
R-HSA-1643685 Disease 3 R-HSA-392499 Metabolism of proteins 3 R-HSA-74160 Gene expression (Transcription) 3 R-HSA-168256 Immune System 2 R-HSA-9612973 Autophagy 2
Partners
NFE2L1PDCD1RAD23
Complex memberships
Png1-Rad23 complex

Evidence

Reading pass · 27 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 PNG1 (yeast ortholog of NGLY1) encodes a soluble cytosolic/nuclear peptide:N-glycanase; when expressed in E. coli it exhibits PNGase enzymatic activity, cleaving N-glycans from glycoproteins/glycopeptides. Subcellular fractionation showed Png1p is present in both nucleus and cytosol. Loss of png1 function reduces efficiency of proteasome-mediated degradation of a misfolded glycoprotein. Genetic screen for PNGase-deficient mutants, recombinant expression in E. coli with enzymatic activity assay, subcellular fractionation/localization The Journal of cell biology High 10831608
2001 Yeast Png1p (NGLY1 ortholog) physically interacts with Rad23p, a ubiquitin-binding protein that links substrates to the 26S proteasome. The Png1p-Rad23p complex is distinct from the Rad4p-Rad23p DNA repair complex. Rad23p is proposed to act as an escort linking Png1p (and thus deglycosylated substrates) to the 26S proteasome. Two-hybrid screening, co-immunoprecipitation (in vivo biochemical confirmation) The Journal of biological chemistry High 11259433
2006 The Png1-Rad23 complex in yeast is required for efficient degradation of a glycosylated ERAD substrate (glycosylated ricin A chain), coupling protein deglycosylation to proteasomal degradation. Rad23 binds various regulators of proteolysis to facilitate degradation of distinct substrates, with substrate specificity determined by interactions with cofactors. Genetic deletion of PNG1/RAD23, glycoprotein turnover assay (glycosylated ricin A chain degradation), protein interaction studies The Journal of cell biology High 16401726
2009 The Neurospora crassa PNG1 ortholog has substitutions in essential catalytic amino acids, abolishing deglycosylation activity, yet PNG1 is essential for cell polarity; its deletion causes strong polarity defects. This reveals an enzyme-independent (non-catalytic) scaffolding function of PNG1 in polar cell growth, distinct from its role in ERAD. Gene deletion, catalytic mutant analysis, morphological phenotyping in Neurospora crassa The Journal of biological chemistry Medium 19940117
2010 Loss-of-function mutations in C. elegans png-1 (NGLY1 ortholog) cause increased axon branching during morphogenesis of vulval egg-laying neurons (VC4, VC5) and nearby axons. PNG-1 acts from both neurons and epithelial cells to restrict axon branching. Genetic interaction with rad-23 (Rad23 ortholog) shows similar branching defects, placing png-1 and rad-23 in the same pathway regulating neuronal branching during organ innervation. Loss-of-function genetic screen in C. elegans, neuronal morphology analysis, tissue-specific rescue, genetic epistasis with rad-23 The Journal of neuroscience High 20130186
2015 In Ngly1-/- mouse embryonic fibroblasts (MEFs), loss of Ngly1 causes delayed degradation of misfolded ERAD substrates. In the absence of Ngly1, ENGase (endo-β-N-acetylglucosaminidase) performs an alternative deglycosylation reaction generating N-GlcNAc proteins that are aggregation-prone. Additional knockout of ENGase in Ngly1-/- cells restores normal ERAD processing, demonstrating that ENGase-generated N-GlcNAc protein aggregates underlie ERAD dysregulation in NGLY1 deficiency. MEF knockout cell lines (Ngly1-/- and Ngly1-/-/ENGase-/-), glycoprotein degradation assays, protein aggregation analysis, genetic epistasis Proceedings of the National Academy of Sciences of the United States of America High 25605922
2017 NGLY1 (human cytosolic N-glycanase) is essential for activation of the transcription factor NFE2L1 (Nrf1) in response to proteasome inhibition. NGLY1 processes retrotranslocated, N-glycosylated Nrf1 by removing its N-glycans; chemical or genetic disruption of NGLY1 results in misprocessed Nrf1 that is excluded from the nucleus and cannot upregulate proteasome subunit gene expression (the 'bounce-back' response). A small-molecule NGLY1 inhibitor was identified that disrupts Nrf1 processing and potentiates proteasome inhibitor cytotoxicity. Genetic KO, siRNA knockdown, small-molecule NGLY1 inhibitor, reporter assays for Nrf1 nuclear localization and transcriptional activity, cell viability assays ACS central science High 29202016
2017 Lethality of Ngly1-/- mice (C57BL/6 background) is partially rescued by additional deletion of the ENGase gene, establishing a genetic epistasis relationship. This demonstrates that ENGase-generated N-GlcNAc protein aggregates are a primary pathological driver in NGLY1 deficiency, and identifies cytoplasmic ENGase as a potential therapeutic target. Double knockout mouse genetics (Ngly1-/- x ENGase-/-), survival analysis, phenotypic rescue PLoS genetics High 28426790
2018 NGLY1 regulates mitochondrial homeostasis through the transcription factor NRF1 (NFE2L1). NGLY1-deficient human and mouse cells show impaired mitochondrial clearance by mitophagy, resulting in fragmented mitochondria and chronic activation of cytosolic nucleic acid-sensing pathways (cGAS-STING and MDA5-MAVS), leading to elevated interferon gene signature. Pharmacological activation of NRF2, a non-glycosylated homolog of NRF1, restores mitochondrial homeostasis and suppresses immune gene activation in NGLY1-deficient cells. NGLY1-deficient human and mouse cell lines, mitophagy assays, mitochondrial morphology/function assays, cGAS-STING/MDA5-MAVS pathway activation measurements, NRF2 activator rescue experiments The Journal of experimental medicine High 30135079
2018 In a Drosophila model of NGLY1 deficiency (loss of Pngl), transcriptome analysis shows no evidence of ER stress but reveals strong NRF1 dysfunction signature (downregulation of proteasome components and oxidation-reduction genes). Loss of NGLY1 is functionally linked to defects in neuroendocrine signaling; targeted NGLY1 expression in prothoracic gland (ecdysteroid-producing tissue) or supplementation with the molting hormone 20-hydroxyecdysone partially rescues developmental delay. Drosophila Pngl loss-of-function model, RNAseq transcriptome analysis, tissue-specific rescue (Gal4/UAS), pharmacological rescue (20-hydroxyecdysone) Human molecular genetics Medium 29346549 29735526
2020 Loss of NGLY1 in Drosophila visceral muscle leads to severely reduced AMPKα (AMP-activated protein kinase α) levels, causing energy metabolism defects, impaired gut peristalsis, and animal lethality. Reduced AMPKα levels are also observed in Ngly1-/- mouse embryonic fibroblasts and NGLY1-deficient patient fibroblasts. Pharmacological AMPK activation suppresses energy metabolism defects. This AMPKα reduction is not caused by loss of NFE2L1 activity, defining an NRF1-independent NGLY1 pathway. Drosophila tissue-specific genetics, Ngly1-/- MEFs, patient fibroblasts, AMPKα western blot, pharmacological AMPK activation, epistasis with NFE2L1 PLoS genetics High 33315951
2020 Loss of NGLY1 in a Drosophila genetic diversity panel reveals NKCC1/2 (ion cotransporter; Drosophila Ncc69) as a modifier of NGLY1 deficiency lethality. In NGLY1-/- mouse cells, NKCC1 shows altered average molecular weight and reduced function, suggesting NKCC1 is a relevant NGLY1 substrate or functionally dependent on NGLY1 activity. Drosophila NGLY1 deficiency model crossed to genetically diverse strains, association analysis, evolutionary rate covariation, NGLY1-/- mouse cell NKCC1 functional assays eLife Medium 33315011
2014 Yeast Png1p deglycosylates ricin A chain (RTA) in the cytosol, and this deglycosylation increases RTA depurination activity by apparently protecting it from ERAD-mediated degradation. In contrast, for a less toxic G83D variant, Png1p deglycosylation on the ER membrane promotes its degradation and reduces toxicity, demonstrating that the free cytosolic pool vs. ER-membrane-associated Png1 have distinct substrate preferences and opposing functional consequences. Yeast genetic deletion of PNG1, EGFP-tagged RTA trafficking assays, depurination activity assays, toxicity assays PloS one Medium 25436896
2019 Loss of Ngly1 in hepatocyte-specific conditional knockout mice causes impaired processing and nuclear localization of Nfe2l1 (NRF1) in hepatocytes, contributing to abnormal hepatocyte nuclear morphology and lipid accumulation under high-fructose diet stress. This establishes that NGLY1 is required for proper Nfe2l1 processing and function in liver tissue in vivo. Liver-specific Cre-loxP conditional Ngly1 knockout mice, Nfe2l1 processing/localization assays, liver phenotype analysis under dietary stress Biochimica et biophysica acta. Molecular basis of disease Medium 31733337
2022 NFE2L1/NRF1 promotes ferroptosis resistance through maintaining expression of glutathione peroxidase 4 (GPX4), and this function requires NGLY1-dependent processing of NFE2L1. NGLY1-mediated NFE2L1 activation is independent of NFE2L2/NRF2, establishing that the NGLY1-NFE2L1 axis constitutes a distinct pathway regulating ferroptosis. NGLY1/NFE2L1 genetic KO cell lines, ferroptosis assays, GPX4 expression analysis, epistasis between NFE2L1 and NFE2L2 Proceedings of the National Academy of Sciences of the United States of America Medium 35271393
2022 In C. elegans, PNG-1/NGLY1 deglycosylates the transcription factor SKN-1A/Nrf1 by converting N-glycosylated asparagine residues to aspartate ('sequence editing'), and this chemical conversion is strictly required for SKN-1A transcriptional activation of proteasome subunit genes. Sequence-edited SKN-1A can also activate redox homeostasis and xenobiotic detoxification genes normally regulated by SKN-1C/Nrf2, but sequence editing itself antagonizes the extent of this activation. C. elegans genetics, transcriptomic analysis, mutant alleles selectively inactivating SKN-1A or SKN-1C, deglycosylation/sequence editing assays bioRxivpreprint Medium
2022 NGLY1 inhibition (pharmacological via Z-VAD-fmk, or siRNA knockdown) induces upregulation of autophagosome formation without impairing autophagic flux in HEK293 cells. This autophagy induction does not involve ER stress markers, ROS, or altered Ca2+ handling. ATG13-deficient MEFs show reduced viability under NGLY1 inhibition, establishing that autophagy is a cytoprotective adaptation to NGLY1 loss. Pharmacological NGLY1 inhibition (Z-VAD-fmk), siRNA knockdown, GFP-LC3 puncta assay, autophagosome immunoprecipitation with mass spectrometry, ATG13-KO MEFs The FEBS journal Medium 34995415
2024 NGLY1 physically interacts with and deglycosylates cancer cell-intrinsic PD-1 in response to doxorubicin treatment. Doxorubicin promotes the interaction between NGLY1 and PD-1, facilitating NGLY1-mediated PD-1 deglycosylation and destabilization (shortening PD-1 half-life), thereby sensitizing tumor cells to doxorubicin's antiproliferative effects. Co-immunoprecipitation (NGLY1-PD-1 interaction), protein half-life assay, siRNA knockdown of NGLY1, doxorubicin treatment, cell viability assays FEBS letters Medium 38782868
2024 Crystal structure of bovine FBS2 (component of SCF-FBS2 ubiquitin ligase) complexed with SKP1 and the N-glycan core pentasaccharide Man3GlcNAc2 was determined, revealing the structural basis for sugar recognition. NMR data revealed disparate sugar-binding specificities among homologous FBS proteins and identified a druggable pocket. FBS2 recognizes N-glycan remnants on proteins processed in the NGLY1/ERAD pathway. X-ray crystallography, NMR spectroscopy, in silico docking FEBS letters High 39171510
2024 NGLY1 possesses a 'sequence editing' enzymatic function: it converts N-glycosylated asparagine residues to aspartate residues on substrate proteins (not merely removing glycans), introducing negative charges into the core peptide. An ELISA-based assay using this Asn-to-Asp conversion to generate a neo-epitope confirmed endogenous NGLY1 activity detectable in as few as 5×10³ cells, including patient-derived iPSCs. ELISA-based activity assay exploiting Asn-to-Asp conversion (neo-epitope detection by anti-HA antibody), crude cell extract from patient and control iPSCs Biochemical and biophysical research communications Medium 38581946
2024 A FRET-based probe assay revealed significant changes in NGLY1 enzymatic activity in rat brains during aging, demonstrating that endogenous NGLY1 activity is dynamically regulated in the central nervous system under physiological conditions. FRET-based fluorescent glycopeptide probe, plate-based assay of endogenous NGLY1 activity in rat brain tissues across ages The Journal of biological chemistry Medium 38417795
2025 The STING pathway drives noninflammatory neurodegeneration in NGLY1 deficiency. In postnatal inducible whole-body Ngly1 knockout mice, genetic ablation of Sting1 rescues Purkinje cell loss, improves motor function, and extends lifespan without requiring immune/inflammatory activation. Single-nucleus RNA sequencing reveals STING-dependent suppression of cholesterol biosynthesis in glia. Pharmacological STING inhibition with VS-X4 mitigates neuropathology and motor disease. Postnatal inducible Ngly1 KO mouse, genetic Sting1 ablation (double KO), single-nucleus RNA sequencing, pharmacological STING inhibition (VS-X4), motor function assays, histopathology The Journal of experimental medicine High 40644312
2023 Loss of Drosophila N-glycanase 1 (Pngl) in a specific intestinal cell type causes gut barrier defects leading to starvation and JNK overactivation; combined loss in enterocytes and fat body causes Foxo overactivation, hyperactive innate immune response, and enhanced lipid catabolism contributing to lethality. Germ-free rearing rescues developmental delay but not lethality, indicating lethality is primarily driven by non-bacterial immune/metabolic mechanisms rather than microbiota. Tissue-specific Drosophila Pngl knockouts, germ-free rearing, fat-rich diet rescue, JNK/Foxo pathway analysis, intestinal barrier assay Nature communications High 37704604
2023 NGLY1 deficiency in human iPSC-derived cortical neurons (but not astrocytes) causes protein aggregate accumulation, mitochondrial homeostasis defects, and synaptic dysfunction. Laser capture microscopy and mass spectrometry characterized the composition of protein aggregates specific to NGLY1-deficient neurons. Introduction of functional NGLY1 rescued these phenotypes, confirming direct causality. iPSC direct conversion to cortical neurons, transcriptomics, proteomics, functional assays, NGLY1 rescue, laser capture microscopy and mass spectrometry of aggregates Cell reports High 38039131
2024 Mutations in nucleotide metabolism genes (rsks-1, tald-1, ent-4) suppress proteasome inhibitor sensitivity caused by PNG-1/NGLY1 inactivation in C. elegans through an SKN-1/Nrf1-independent mechanism. Restriction of nucleotide availability (via ent-4 intestinal nucleoside/nucleotide transporter mutation or dietary manipulation) is beneficial, while a nucleotide-rich diet exacerbates proteasome dysfunction in PNG-1/NGLY1-deficient worms. C. elegans suppressor screen, double mutant epistasis, dietary nucleotide manipulation, proteasome function assays PLoS biology Medium 38991033
2025 Natural variants in SEL1L (cytoplasmic tail domain, S780P and Δ806-809) modify NGLY1 deficiency lethality in Drosophila by enhancing ERAD function in an NGLY1-dependent manner. This genetic interaction implicates NGLY1 as a contributor to general ERAD function (not solely deglycosylation of specific substrates), and SEL1L variants also protect against proteasome inhibition sensitivity in heterozygous NGLY1 null flies. CRISPR-generated Drosophila SEL1L variant lines, NGLY1 deficiency model genetic interaction, ERAD functional assays, ER stress resistance assays, proteasome inhibition assays PLoS genetics Medium 40773511
2025 NGLY1 exhibits a novel exo-(N-)glycanase activity on ENGase-digested N-GlcNAc proteins in vitro, distinct from its canonical endo-(N-)glycanase activity. Active sites for exo-NGLY1 activity were predicted computationally and differ from those of endo-NGLY1. In vitro enzymatic assay with N-GlcNAc protein substrates, computational active-site prediction/comparison bioRxivpreprint Low

Source papers

Stage 0 corpus · 97 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 Mutations in NGLY1 cause an inherited disorder of the endoplasmic reticulum-associated degradation pathway. Genetics in medicine : official journal of the American College of Medical Genetics 181 24651605
2000 PNG1, a yeast gene encoding a highly conserved peptide:N-glycanase. The Journal of cell biology 179 10831608
2017 Inhibition of NGLY1 Inactivates the Transcription Factor Nrf1 and Potentiates Proteasome Inhibitor Cytotoxicity. ACS central science 156 29202016
2016 Prospective phenotyping of NGLY1-CDDG, the first congenital disorder of deglycosylation. Genetics in medicine : official journal of the American College of Medical Genetics 124 27388694
2006 The Png1-Rad23 complex regulates glycoprotein turnover. The Journal of cell biology 108 16401726
2018 N-glycanase NGLY1 regulates mitochondrial homeostasis and inflammation through NRF1. The Journal of experimental medicine 106 30135079
2015 Endo-β-N-acetylglucosaminidase forms N-GlcNAc protein aggregates during ER-associated degradation in Ngly1-defective cells. Proceedings of the National Academy of Sciences of the United States of America 105 25605922
2001 Rad23 provides a link between the Png1 deglycosylating enzyme and the 26 S proteasome in yeast. The Journal of biological chemistry 105 11259433
2015 The cytoplasmic peptide:N-glycanase (NGLY1) - Structure, expression and cellular functions. Gene 104 26611529
2022 Ferroptosis regulation by the NGLY1/NFE2L1 pathway. Proceedings of the National Academy of Sciences of the United States of America 85 35271393
2017 Lethality of mice bearing a knockout of the Ngly1-gene is partially rescued by the additional deletion of the Engase gene. PLoS genetics 81 28426790
2014 NGLY1 mutation causes neuromotor impairment, intellectual disability, and neuropathy. European journal of medical genetics 72 25220016
2014 The cytoplasmic peptide:N-glycanase (Ngly1)-basic science encounters a human genetic disorder. Journal of biochemistry 55 25398991
2020 Ngly1 -/- rats develop neurodegenerative phenotypes and pathological abnormalities in their peripheral and central nervous systems. Human molecular genetics 53 32259258
2015 Multi-systemic involvement in NGLY1-related disorder caused by two novel mutations. American journal of medical genetics. Part A 51 25707956
2017 Mitochondrial function requires NGLY1. Mitochondrion 49 28750948
2018 Transcriptome and functional analysis in a Drosophila model of NGLY1 deficiency provides insight into therapeutic approaches. Human molecular genetics 47 29346549
2009 The Neurospora peptide:N-glycanase ortholog PNG1 is essential for cell polarity despite its lack of enzymatic activity. The Journal of biological chemistry 42 19940117
1997 Png-1, a nervous system-specific zinc finger gene, identifies regions containing postmitotic neurons during mammalian embryonic development. The Journal of comparative neurology 42 9130664
2003 Ngly1, a mouse gene encoding a deglycosylating enzyme implicated in proteasomal degradation: expression, genomic organization, and chromosomal mapping. Biochemical and biophysical research communications 40 12711318
2019 Aspartylglycosamine is a biomarker for NGLY1-CDDG, a congenital disorder of deglycosylation. Molecular genetics and metabolism 39 31311714
2010 The N-glycanase png-1 acts to limit axon branching during organ formation in Caenorhabditis elegans. The Journal of neuroscience : the official journal of the Society for Neuroscience 38 20130186
2017 Repurposing of Proton Pump Inhibitors as first identified small molecule inhibitors of endo-β-N-acetylglucosaminidase (ENGase) for the treatment of NGLY1 deficiency, a rare genetic disease. Bioorganic & medicinal chemistry letters 32 28512024
2020 Variants in NGLY1 lead to intellectual disability, myoclonus epilepsy, sensorimotor axonal polyneuropathy and mitochondrial dysfunction. Clinical genetics 31 31957011
2020 A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency. eLife 31 33315011
2020 A conserved role for AMP-activated protein kinase in NGLY1 deficiency. PLoS genetics 31 33315951
2022 NGLY1 Deficiency, a Congenital Disorder of Deglycosylation: From Disease Gene Function to Pathophysiology. Cells 30 35406718
2019 Unexplained death in patients with NGLY1 mutations may be explained by adrenal insufficiency. Physiological reports 29 30740912
2020 NGLY1 deficiency-A rare congenital disorder of deglycosylation. JIMD reports 27 32395402
2021 Reversibility of motor dysfunction in the rat model of NGLY1 deficiency. Molecular brain 25 34120625
2020 Novel NGLY1 gene variants in Chinese children with global developmental delay, microcephaly, hypotonia, hypertransaminasemia, alacrimia, and feeding difficulty. Journal of human genetics 25 31965062
2019 Liver-specific deletion of Ngly1 causes abnormal nuclear morphology and lipid metabolism under food stress. Biochimica et biophysica acta. Molecular basis of disease 25 31733337
2021 NGLY1 deficiency: Novel variants and literature review. European journal of medical genetics 24 33497766
2021 JF1/B6F1 Ngly1-/- mouse as an isogenic animal model of NGLY1 deficiency. Proceedings of the Japan Academy. Series B, Physical and biological sciences 24 33563880
2019 Drug screens of NGLY1 deficiency in worm and fly models reveal catecholamine, NRF2 and anti-inflammatory-pathway activation as potential clinical approaches. Disease models & mechanisms 24 31615832
2018 Defects in the Neuroendocrine Axis Contribute to Global Development Delay in a Drosophila Model of NGLY1 Deficiency. G3 (Bethesda, Md.) 24 29735526
2022 GlcNAc-Asn is a biomarker for NGLY1 deficiency. Journal of biochemistry 22 34697629
2022 N-glycoproteomics reveals distinct glycosylation alterations in NGLY1-deficient patient-derived dermal fibroblasts. Journal of inherited metabolic disease 22 36102038
2022 AAV9-NGLY1 gene replacement therapy improves phenotypic and biomarker endpoints in a rat model of NGLY1 Deficiency. Molecular therapy. Methods & clinical development 20 36320418
2019 Transiently elevated plasma methionine, S-adenosylmethionine and S-adenosylhomocysteine: Unreported laboratory findings in a patient with NGLY1 deficiency, a congenital disorder of deglycosylation. JIMD reports 20 31497478
2020 Acute liver failure in a male patient with NGLY1-congenital disorder of deglycosylation. European journal of medical genetics 19 32422350
2023 Gut barrier defects, intestinal immune hyperactivation and enhanced lipid catabolism drive lethality in NGLY1-deficient Drosophila. Nature communications 18 37704604
2021 NGLY1 Deficiency: A Rare Newly Described Condition with a Typical Presentation. Life (Basel, Switzerland) 18 33673403
2018 Stress and interferon signalling-mediated apoptosis contributes to pleiotropic anticancer responses induced by targeting NGLY1. British journal of cancer 18 30385822
2022 An in vivo drug repurposing screen and transcriptional analyses reveals the serotonin pathway and GSK3 as major therapeutic targets for NGLY1 deficiency. PLoS genetics 17 35653343
2020 NGLY1 deficiency: Novel patient, review of the literature and diagnostic algorithm. JIMD reports 17 32071843
2023 Generation and characterization of NGLY1 patient-derived midbrain organoids. Frontiers in cell and developmental biology 16 36875753
2023 NGLY1 mutations cause protein aggregation in human neurons. Cell reports 16 38039131
2022 Delineating the epilepsy phenotype of NGLY1 deficiency. Journal of inherited metabolic disease 16 35243670
2022 Off-target inhibition of NGLY1 by the polycaspase inhibitor Z-VAD-fmk induces cellular autophagy. The FEBS journal 15 34995415
2023 NGLY1 deficiency: a prospective natural history study. Human molecular genetics 14 37379343
2023 NGLY1: A fascinating, multifunctional molecule. Biochimica et biophysica acta. General subjects 14 37951368
2022 Physiological importance of NGLY1, as revealed by rodent model analyses. Journal of biochemistry 12 34580715
2022 Assay for the peptide:N-glycanase/NGLY1 and disease-specific biomarkers for diagnosing NGLY1 deficiency. Journal of biochemistry 10 34791337
2022 NGLY1 Deficiency Zebrafish Model Manifests Abnormalities of the Nervous and Musculoskeletal Systems. Frontiers in cell and developmental biology 10 35769264
2024 Development of a fluorescence and quencher-based FRET assay for detection of endogenous peptide:N-glycanase/NGLY1 activity. The Journal of biological chemistry 9 38417795
2022 NGLY1: insights from Caenorhabditis elegans. Journal of biochemistry 9 34697631
2022 Ever-expanding NGLY1 biology. Journal of biochemistry 9 34969094
2025 The STING pathway drives noninflammatory neurodegeneration in NGLY1 deficiency. The Journal of experimental medicine 7 40644312
2010 Characterization of the temperature-sensitive mutations un-7 and png-1 in Neurospora crassa. PloS one 7 20502699
2022 Cytosolic O-GlcNAcylation and PNG1 maintain Drosophila gut homeostasis by regulating proliferation and apoptosis. PLoS genetics 6 35294432
2018 Generation of an induced pluripotent stem cell line (TRNDi002-B) from a patient carrying compound heterozygous p.Q208X and p.G310G mutations in the NGLY1 gene. Stem cell research 6 30612078
2024 ELISA-based highly sensitive assay system for the detection of endogenous NGLY1 activity. Biochemical and biophysical research communications 5 38581946
2024 Ocular features of NGLY1 deficiency from a prospective longitudinal cohort. Journal of AAPOS : the official publication of the American Association for Pediatric Ophthalmology and Strabismus 5 38697387
2024 Mutations in nucleotide metabolism genes bypass proteasome defects in png-1/NGLY1-deficient Caenorhabditis elegans. PLoS biology 5 38991033
2024 Systemic gene therapy corrects the neurological phenotype in a mouse model of NGLY1 deficiency. JCI insight 5 39137042
2024 Development of new NGLY1 assay systems - toward developing an early screening method for NGLY1 deficiency. Glycobiology 5 39206713
2015 New perspectives on the mutated NGLY1 enigma. Medical hypotheses 5 26228302
2014 Wild type RTA and less toxic variants have distinct requirements for Png1 for their depurination activity and toxicity in Saccharomyces cerevisiae. PloS one 5 25436896
2024 Intranasal oxytocin suppresses seizure-like behaviors in a mouse model of NGLY1 deficiency. Communications biology 4 38649481
2024 Doxorubicin induces deglycosylation of cancer cell-intrinsic PD-1 by NGLY1. FEBS letters 4 38782868
2019 An induced pluripotent stem cell line (TRNDi010-C) from a patient carrying a homozygous p.R401X mutation in the NGLY1 gene. Stem cell research 4 31326749
2024 Structural basis of sugar recognition by SCFFBS2 ubiquitin ligase involved in NGLY1 deficiency. FEBS letters 3 39171510
2022 Comparative proteomics reveals elevated CCN2 in NGLY1-deficient cells. Biochemical and biophysical research communications 3 36209585
2022 An induced pluripotent stem cell-derived NMJ platform for study of the NGLY1-Congenital Disorder of Deglycosylation. Advanced therapeutics 3 36589922
2021 Generation of two gene corrected human isogenic iPSC lines (NCATS-CL6104 and NCATS-CL6105) from a patient line (NCATS-CL6103) carrying a homozygous p.R401X mutation in the NGLY1 gene using CRISPR/Cas9. Stem cell research 3 34619643
2025 Increased oxidative stress and autophagy in NGLY1 patient iPSC-derived neural stem cells. Experimental cell research 2 40189184
2025 Impaired Proteostasis is Linked to Neurological Pathology in a Zebrafish NGLY1 Deficiency Model. Journal of inherited metabolic disease 2 40470739
2025 NGLY1 deficiency - clinical features and therapeutic strategy. Journal of human genetics 2 40730667
2025 Natural SEL1L variants rescue a model of NGLY1 deficiency and modify ERAD function and proteasome sensitivity. PLoS genetics 2 40773511
2025 Tofacitinib Improves Motor Symptoms in Parkinsonism Associated with a Heterozygous NGLY1 Variant and Autoimmune Disease. European journal of case reports in internal medicine 2 41229635
2024 Functional prediction of the potential NGLY1 mutations associated with rare disease CDG. Heliyon 2 38628705
2025 An Assay System for Plate-based Detection of Endogenous Peptide:N-glycanase/NGLY1 Activity Using A Fluorescence-based Probe. Bio-protocol 1 39803314
2025 Preclinical pharmacology and safety studies to support an AAV9 NGLY1 gene therapy clinical trial for the treatment of NGLY1 deficiency. Molecular therapy. Methods & clinical development 1 40687377
2025 Comparative proteomics of HepG2 cells reveals NGLY1 as an important regulator of ferroptosis resistance and iron uptake. PloS one 1 40811347
2025 AAV9-mediated NGLY1 gene replacement suppresses non-epileptic convulsions in Ngly1-/- rats. Biochemical and biophysical research communications 1 41176936
2023 A Natural Compound Containing a Disaccharide Structure of Glucose and Rhamnose Identified as Potential N-Glycanase 1 (NGLY1) Inhibitors. Molecules (Basel, Switzerland) 1 38067490
2021 An induced pluripotent stem cell line (NCATS-CL9075) from a patient carrying compound heterozygote mutations, p.R390P and p.L318P, in the NGLY1 gene. Stem cell research 1 34051448
2026 Progressive neurodegeneration, motor decline, and premature mortality in aging Ngly1 deficient rats. Orphanet journal of rare diseases 0 41721346
2026 Cytosolic Peptide: N-Glycanase (NGLY1)-from Basic Biology to Genetic Disorder, NGLY1 Deficiency. Advances in experimental medicine and biology 0 41917400
2026 Discovering the Hidden Power of NGLY1: Orchestrating Immune Cell Functions and Autoimmune Diseases. Frontiers in bioscience (Landmark edition) 0 42052850
2026 Natural history of NGLY1 deficiency: motor function & clinical features. Human molecular genetics 0 42114141
2025 Unveiling roles of NGLY1 in cellular homeostasis and related diseases. Cellular signalling 0 40602583
2025 NGLY1-CDDG: report of two cases from India and brief review of literature. Journal of genetics 0 40810208
2025 Structural characterization and insights into the formation of N-acetylglucosaminylasparagine and its derivatives in NGLY1-deficient models and patients. Glycobiology 0 41081738
2025 NGLY1 as an Emerging Critical Modulator for Neurodevelopment and Pathogenesis in the Brain. International journal of molecular sciences 0 41096971
2023 Sugar coating autophagy: exploring the links between the inhibition of NGLY1 (N-glycanase 1) and autophagy induction. Autophagy reports 0 40395312

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