Affinage

NT5DC2

5'-nucleotidase domain-containing protein 2 · UniProt Q9H857

Length
520 aa
Mass
60.7 kDa
Annotated
2026-06-10
17 papers in source corpus 14 papers cited in narrative 14 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NT5DC2 is a cytoplasmic HAD-phosphatase-domain protein that functions both as a regulator of catecholamine-metabolizing enzymes and as a pro-survival, pro-proliferative factor in multiple cancers by stabilizing client proteins against ubiquitin-proteasome degradation (PMID:38382359, PMID:32382041). In catecholaminergic cells, NT5DC2 binds tyrosine hydroxylase and acts as (or recruits) a phosphatase that dephosphorylates TH at Ser40, suppressing DOPA and catecholamine synthesis; purified NT5DC2 binds phosphorylated recombinant TH and promotes its dephosphorylation in vitro, while it preferentially associates with non-phosphorylated MAO A to enhance MAO A activity and dopamine metabolism (PMID:31279527, PMID:32778969, PMID:38382359, PMID:40751758). In tumor cells, NT5DC2 directly binds EGFR and reduces its ubiquitination to prevent proteasomal degradation, sustaining downstream EGFR signaling and glycolytic metabolism in hepatocellular carcinoma and triple-negative breast cancer (PMID:32382041, PMID:38289126). This stabilization role extends to additional clients: NT5DC2 protects unpalmitoylated TEAD4 from TRIM27-mediated ubiquitination (with TEAD4 reciprocally activating the NT5DC2 promoter in a feedback loop) and stabilizes the ferroptosis suppressor ACSL3 by inhibiting its ubiquitination, thereby restraining ferroptosis (PMID:33993634, PMID:41974665). Through these activities NT5DC2 supports proliferation and survival across cancer types, acting upstream of Fyn in glioma stem-like cells, upstream of p53-dependent apoptosis and G2 arrest in NSCLC, and within a HIF-1α/VEGF-A/CCL2 angiogenic axis in colorectal cancer (PMID:30978441, PMID:32962856, PMID:32991874). NT5DC2 expression is itself controlled post-transcriptionally and transcriptionally: the RNA-binding protein IGF2BP2 and the helicase DDX3X bind NT5DC2 mRNA to promote its expression/translation in DLBCL and CML, and CTCF (stabilized via IGF2BP3/m6A) transcriptionally activates NT5DC2 in lung squamous cell carcinoma (PMID:35894142, PMID:39516658, PMID:39506204).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 2019 Medium

    Establishing the first molecular partner of NT5DC2 showed it physically engages tyrosine hydroxylase and negatively regulates catecholamine output without altering TH abundance, implying a catalytic rather than stabilizing effect.

    Evidence Anti-TH immunoprecipitation with nano-LC-MS/MS and siRNA knockdown with catecholamine measurement in PC12D cells

    PMID:31279527

    Open questions at the time
    • Direct enzymatic mechanism on TH not yet demonstrated
    • Phosphorylation site not yet defined
    • Single cell type
  2. 2020 Medium

    Mapping NT5DC2's effect to TH Ser40 dephosphorylation defined the regulatory readout linking NT5DC2 to DOPA synthesis control.

    Evidence siRNA knockdown, phospho-Ser40 Western blot, DOPA measurement and immunocytochemistry co-localization

    PMID:32778969

    Open questions at the time
    • Whether NT5DC2 is the catalytic phosphatase or an activator unresolved
    • No in vitro reconstitution at this stage
  3. 2020 High

    Identifying EGFR as a direct binding partner that NT5DC2 stabilizes against ubiquitin-proteasome degradation established its first oncogenic stabilization mechanism and a pharmacologically testable pathway.

    Evidence Co-IP/LC-MS/MS, ubiquitination assay, erlotinib rescue, and proliferation assays in hepatocellular carcinoma

    PMID:32382041

    Open questions at the time
    • Whether stabilization requires phosphatase activity unknown
    • Direct binding interface not mapped
  4. 2020 Medium

    Placing NT5DC2 upstream of p53 and within a HIF-1α/VEGF-A/CCL2 axis extended its pro-survival role to apoptosis suppression and angiogenesis in additional cancers.

    Evidence siRNA/shRNA knockdown with epistasis (NT5DC2+p53; VEGF rescue), flow cytometry, tube formation, and xenograft models in NSCLC and CRC

    PMID:32962856 PMID:32991874

    Open questions at the time
    • Direct molecular link between NT5DC2 and p53 or HIF-1α not defined
    • Mechanism may be indirect via EGFR
  5. 2019 Medium

    Demonstrating that NT5DC2 knockdown reduces Fyn and impairs glioma stem-cell maintenance positioned NT5DC2 as a stemness regulator upstream of a Src-family kinase.

    Evidence siRNA/shRNA knockdown, tumorsphere and viability assays, xenograft, Fyn Western blot in glioma stem-like cells

    PMID:30978441

    Open questions at the time
    • Whether Fyn regulation is direct or transcriptional unknown
    • No physical interaction shown
  6. 2021 Medium

    Showing NT5DC2 shields unpalmitoylated TEAD4 from TRIM27-mediated ubiquitination, with TEAD4 reactivating the NT5DC2 promoter, revealed a self-reinforcing stabilization circuit.

    Evidence Co-IP, ubiquitination site mapping (Lys278), dual-luciferase reporter, and proliferation assays

    PMID:33993634

    Open questions at the time
    • Whether NT5DC2 competes directly with TRIM27 unknown
    • Role of NT5DC2 phosphatase activity in protection untested
  7. 2022 Medium

    Identifying IGF2BP2 as an RNA-binding regulator of NT5DC2 mRNA established post-transcriptional control of NT5DC2 levels driving the survival phenotype.

    Evidence RNA pulldown, RIP, knockdown/overexpression epistasis, flow cytometry in DLBCL

    PMID:35894142

    Open questions at the time
    • m6A dependence of IGF2BP2 binding not tested here
    • Effect on translation vs stability not separated
  8. 2024 High

    In vitro reconstitution with purified proteins demonstrated that NT5DC2 binds phosphorylated TH and directly promotes its dephosphorylation, advancing the model from correlation to a phosphatase-like activity.

    Evidence In vitro binding and dephosphorylation assays with purified recombinant NT5DC2 and rhTH1, overexpression DOPA measurement

    PMID:38382359

    Open questions at the time
    • Whether NT5DC2 itself is catalytic or an obligate cofactor still not fully resolved
    • Active-site residues not mutated
  9. 2024 Medium

    Extending the EGFR-stabilization mechanism to TNBC and linking it to glycolytic metabolism generalized NT5DC2's role in EGFR-driven tumor metabolism.

    Evidence Co-IP, siRNA knockdown with EGFR-pathway rescue, Seahorse metabolic assays, xenograft in TNBC

    PMID:38289126

    Open questions at the time
    • Direct binding interface unmapped
    • Metabolic effect may be entirely downstream of EGFR
  10. 2024 Medium

    Defining DDX3X-driven translation and CTCF-driven transcription (via IGF2BP3/m6A) of NT5DC2 placed it within layered upstream regulatory axes governing leukemia stem cells and lung squamous carcinoma.

    Evidence RIP, shRNA knockdown with epistasis rescue (CML model); dual-luciferase, m6A RIP, knockdown/xenograft (LUSC)

    PMID:39506204 PMID:39516658

    Open questions at the time
    • Whether these regulators are tissue-specific or general unknown
    • Direct vs indirect regulation of NT5DC2 not fully separated
  11. 2025 Medium

    Identifying preferential binding to non-phosphorylated MAO A and enhancement of its activity broadened NT5DC2's role in monoamine metabolism beyond TH.

    Evidence AP-MS, MAO A binding Western blot, siRNA knockdown with MAO A activity and metabolite assays in PC12D cells

    PMID:40751758

    Open questions at the time
    • Mechanism by which NT5DC2 enhances MAO A activity undefined
    • Phosphorylation-state dependence of binding not mechanistically explained
  12. 2026 Medium

    Showing NT5DC2 stabilizes ACSL3 by inhibiting its ubiquitination and restrains ferroptosis added a lipid-metabolism/cell-death dimension to its stabilization function.

    Evidence Co-IP, ubiquitination assay, ferroptosis assays, ACSL3 rescue, oleic acid stimulation, xenograft in bladder cancer

    PMID:41974665

    Open questions at the time
    • Responsible E3 ligase not identified
    • Whether stabilization needs phosphatase activity untested

Open questions

Synthesis pass · forward-looking unresolved questions
  • Whether NT5DC2's HAD phosphatase activity is the unifying biochemical mechanism behind both its TH/MAO A regulation and its many ubiquitination-blocking stabilization roles remains unresolved.
  • No catalytic-dead mutant tested across its client proteins
  • No structural model of substrate engagement
  • Connection between catecholamine and cancer functions unestablished

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 3 GO:0140313 molecular sequestering activity 3
Localization
GO:0005829 cytosol 1
Pathway
R-HSA-1430728 Metabolism 3 R-HSA-162582 Signal Transduction 2 R-HSA-5357801 Programmed Cell Death 2
Partners
ACSL3EGFRMAOATEAD4TH

Evidence

Reading pass · 14 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2019 NT5DC2 physically binds to tyrosine hydroxylase (TH) as identified by nano-LC-MS/MS analysis of immunoprecipitates from PC12D cells; siRNA-mediated knockdown of NT5DC2 increased catecholamine (dopamine, noradrenaline, adrenaline) synthesis without changing TH protein levels, indicating NT5DC2 inhibits TH catalytic activity rather than its stability. Immunoprecipitation with anti-TH antibody followed by nano-LC-MS/MS; siRNA knockdown with catecholamine synthesis measurement Biochemical and biophysical research communications Medium 31279527
2020 NT5DC2 decreases DOPA synthesis by promoting dephosphorylation of TH at Ser40; siRNA knockdown of NT5DC2 increased TH phosphorylation at Ser residues and enhanced DOPA synthesis, and immunocytochemistry showed NT5DC2 and TH co-localize in the cytoplasm, with NT5DC2 proposed to act as a phosphatase or phosphatase-activator on TH. siRNA knockdown, Western blot for phospho-TH (Ser40), DOPA/catecholamine measurement, immunocytochemistry for co-localization Journal of neural transmission Medium 32778969
2024 Purified NT5DC2-tag protein binds to the phosphorylated form of recombinant human TH type 1 (rhTH1) in vitro; overexpression of NT5DC2 decreased DOPA levels; incubation of cell lysate or purified NT5DC2-tag with phosphorylated rhTH1 at 37°C decreased TH phosphorylation, demonstrating NT5DC2 promotes TH dephosphorylation acting similarly to a phosphatase. In vitro binding assay with purified recombinant proteins, Western blot for phospho-TH, DOPA measurement by overexpression, proteomic analysis by mass spectrometry Biochemical and biophysical research communications High 38382359
2025 NT5DC2 binds primarily to the non-phosphorylated form of monoamine oxidase A (MAO A); siRNA-mediated NT5DC2 downregulation reduced MAO A activity, decreased dopamine metabolism, and increased noradrenaline synthesis in PC12D cells, indicating NT5DC2 promotes MAO A activity. Affinity purification-mass spectrometry (interaction identification), Western blot for MAO A binding, siRNA knockdown with MAO A activity assay and catecholamine/metabolite measurement Molecular biology reports Medium 40751758
2020 NT5DC2 directly binds EGFR (identified by co-immunoprecipitation and LC-MS/MS) and stabilizes EGFR protein by reducing its ubiquitination and preventing proteasomal degradation, thereby activating downstream EGFR signaling to promote hepatocellular carcinoma cell proliferation. Co-immunoprecipitation, LC-MS/MS, ubiquitination assay, Western blot, in vitro and in vivo proliferation assays, EGFR inhibitor (erlotinib) rescue Cell death & disease High 32382041
2019 NT5DC2 knockdown in glioma stem-like cells (GSCs) markedly reduces Fyn expression (a Src family kinase), inhibits tumorsphere formation and cell viability in vitro, and suppresses tumorigenesis in vivo, placing NT5DC2 upstream of Fyn in GSC maintenance. siRNA/shRNA knockdown, tumorsphere formation assay, cell viability assay, in vivo xenograft model, Western blot for Fyn Cancer letters Medium 30978441
2021 NT5DC2 interacts with unpalmitoylated TEAD4 and protects it from ubiquitin-proteasome degradation; TRIM27 is identified as the E3 ubiquitin ligase mediating K27/K48-linked ubiquitination of unpalmitoylated TEAD4 at Lys278; TEAD4 in turn transcriptionally activates NT5DC2 promoter, forming a positive feedback loop. Co-immunoprecipitation, ubiquitination assay, dual-luciferase reporter assay, shRNA knockdown, in vitro and in vivo proliferation assays Journal of cellular and molecular medicine Medium 33993634
2020 NT5DC2 knockdown in NSCLC cells increases p53 expression and p53-dependent apoptosis and G2 arrest; p53 downregulation abrogates the anti-proliferative and pro-apoptotic effects of NT5DC2 knockdown, placing NT5DC2 upstream of p53 in NSCLC cell survival. siRNA knockdown, overexpression, flow cytometry (cell cycle, apoptosis), Western blot, epistasis by double knockdown (NT5DC2 + p53) Biochemical and biophysical research communications Medium 32962856
2020 NT5DC2 knockdown in CRC cells reduces HIF-1α and VEGF-A expression, inhibits angiogenesis (tube formation), suppresses CCL2/CCR2 expression, and blocks AKT/NF-κB signaling; VEGF reduction is necessary for the anti-proliferative, anti-migratory, and anti-angiogenic effects of NT5DC2 knockdown, defining an NT5DC2/VEGF/CCL2 axis. shRNA knockdown (lentiviral), tube formation assay, conditioned medium co-culture, Western blot, in vivo xenograft and lung metastasis model Experimental cell research Medium 32991874
2024 NT5DC2 interacts with EGFR in TNBC cells to promote downstream EGFR signal transduction; NT5DC2 knockdown suppresses glycolysis (reduced extracellular acidification rate, ATP, lactate, glucose uptake), and EGFR pathway activation counteracts the effects of NT5DC2 knockdown, confirming NT5DC2 acts via EGFR. Co-immunoprecipitation, siRNA knockdown, EGFR pathway rescue experiment, Seahorse/metabolic assays, in vivo xenograft Molecular carcinogenesis Medium 38289126
2022 IGF2BP2 RNA-binding protein binds to NT5DC2 mRNA (confirmed by RNA pulldown and immunoprecipitation); IGF2BP2 upregulation reverses the anti-proliferative, pro-apoptotic, and cell-cycle arrest effects of NT5DC2 knockdown in DLBCL cells, demonstrating IGF2BP2 acts upstream of NT5DC2 to regulate its expression post-transcriptionally. RNA pulldown assay, RNA immunoprecipitation, siRNA/shRNA knockdown, overexpression, flow cytometry, Western blot Molecular medicine reports Medium 35894142
2024 DDX3X RNA helicase binds NT5DC2 mRNA and promotes its translation in CML cells; DDX3X inhibition reduces NT5DC2 protein and eliminates leukemia stem cells, with NT5DC2 acting as a functional mediator downstream of DDX3X in CML LSC maintenance. RNA immunoprecipitation (DDX3X binds NT5DC2 mRNA), shRNA knockdown of DDX3X with NT5DC2 protein measurement, epistasis rescue with NT5DC2 overexpression, CML mouse model Oncogene Medium 39516658
2024 CTCF transcriptionally activates NT5DC2 in lung squamous cell carcinoma cells; IGF2BP3 stabilizes CTCF mRNA via m6A methylation, forming an IGF2BP3/CTCF/NT5DC2 regulatory axis; NT5DC2 knockdown inhibits LUSC cell proliferation, glycolysis, and M2 macrophage polarization, and these effects are rescued by NT5DC2 overexpression. Dual-luciferase reporter assay (CTCF→NT5DC2 promoter), RNA immunoprecipitation, m6A RIP assay, siRNA knockdown, flow cytometry, xenograft model The clinical respiratory journal Medium 39506204
2026 NT5DC2 physically interacts with ACSL3 (a ferroptosis suppressor protein) and inhibits its ubiquitination, thereby stabilizing ACSL3 protein; NT5DC2 knockdown promotes ferroptosis in bladder cancer cells, and this effect is rescued by ACSL3; NT5DC2 also mediates oleic acid-induced upregulation of ACSL3. Co-immunoprecipitation, ubiquitination assay, siRNA knockdown, ferroptosis assay, rescue experiment with ACSL3 overexpression, in vivo xenograft Cell death discovery Medium 41974665

Source papers

Stage 0 corpus · 17 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2020 NT5DC2 promotes tumor cell proliferation by stabilizing EGFR in hepatocellular carcinoma. Cell death & disease 57 32382041
2019 NT5DC2 promotes tumorigenicity of glioma stem-like cells by upregulating fyn. Cancer letters 40 30978441
2020 NT5DC2 knockdown inhibits colorectal carcinoma progression by repressing metastasis, angiogenesis and tumor-associated macrophage recruitment: A mechanism involving VEGF signaling. Experimental cell research 26 32991874
2020 NT5DC2 suppression restrains progression towards metastasis of non-small-cell lung cancer through regulation p53 signaling. Biochemical and biophysical research communications 20 32962856
2021 NT5DC2 promotes leiomyosarcoma tumour cell growth via stabilizing unpalmitoylated TEAD4 and generating a positive feedback loop. Journal of cellular and molecular medicine 18 33993634
2019 Identification by nano-LC-MS/MS of NT5DC2 as a protein binding to tyrosine hydroxylase: Down-regulation of NT5DC2 by siRNA increases catecholamine synthesis in PC12D cells. Biochemical and biophysical research communications 17 31279527
2022 Decreased RNA‑binding protein IGF2BP2 downregulates NT5DC2, which suppresses cell proliferation, and induces cell cycle arrest and apoptosis in diffuse large B‑cell lymphoma cells by regulating the p53 signaling pathway. Molecular medicine reports 10 35894142
2020 NT5DC2 affects the phosphorylation of tyrosine hydroxylase regulating its catalytic activity. Journal of neural transmission (Vienna, Austria : 1996) 10 32778969
2024 NT5DC2 knockdown suppresses progression, glycolysis, and neuropathic pain in triple-negative breast cancer by blocking the EGFR pathway. Molecular carcinogenesis 7 38289126
2024 Role of NT5DC2 in tyrosine hydroxylase phosphorylation based on the analysis of NT5DC2-binding proteins. Biochemical and biophysical research communications 5 38382359
2024 Targeting DDX3X eliminates leukemia stem cells in chronic myeloid leukemia by blocking NT5DC2 mRNA translation. Oncogene 5 39516658
2024 IGF2BP3/CTCF Axis-Dependent NT5DC2 Promotes M2 Macrophage Polarization to Enhance the Malignant Progression of Lung Squamous Cell Carcinomas. The clinical respiratory journal 2 39506204
2023 [Corrigendum] Decreased RNA‑binding protein IGF2BP2 downregulates NT5DC2, which suppresses cell proliferation, and induces cell cycle arrest and apoptosis in diffuse large B‑cell lymphoma cells by regulating the p53 signaling pathway. Molecular medicine reports 2 36799157
2026 NT5DC2 inhibits ferroptosis by stabilizing ACSL3 in bladder cancer. Cell death discovery 0 41974665
2025 circ_0046599 Promotes HCC Progression by Competing with miR-1322 to Enhance NT5DC2 Expression. Journal of Cancer 0 40302808
2025 NT5DC2 downregulation suppresses monoamine oxidase activity and promotes catecholamine synthesis in PC12D cells. Molecular biology reports 0 40751758
2024 Quantitative proteomics and immunohistochemistry uncover NT5DC2 as a diagnostic biomarker for papillary urothelial carcinoma. Heliyon 0 39165948

Missed literature

Know a paper Affinage missed for NT5DC2? Flag it for the maintainers and the community.

No submissions yet.