| 2014 |
GPX4 is directly inhibited by a class of ferroptosis-inducing compounds (e.g., RSL3) identified via chemoproteomics; GPX4 overexpression suppresses and knockdown promotes ferroptotic cell death induced by 12 different ferroptosis inducers, establishing GPX4 as an essential regulator of ferroptosis. |
Chemoproteomics (direct target identification), GPX4 overexpression/knockdown with cell viability readouts, targeted metabolomic profiling |
Cell |
High |
24439385
|
| 2019 |
GPX4 functions as the glutathione-dependent lipid hydroperoxidase that converts lipid hydroperoxides into non-toxic lipid alcohols, thereby suppressing ferroptosis; FSP1 acts in a parallel non-mitochondrial CoQ antioxidant pathway independent of GPX4. |
Synthetic lethal CRISPR-Cas9 screen, biochemical oxidoreductase assays, xenograft tumor models |
Nature |
High |
31634900
|
| 2017 |
Selenocysteine utilization by GPX4 confers exquisite resistance to irreversible overoxidation; cells expressing a cysteine variant of GPX4 (Sec→Cys) are highly sensitive to peroxide-induced ferroptosis, demonstrating that selenolate-based catalysis is specifically required for neuronal survival and prevention of fatal epileptic seizures in vivo. |
Knock-in mouse models (Gpx4cys/cys), in vitro peroxide sensitivity assays, neuronal cell-specific conditional knockouts |
Cell |
High |
29290465
|
| 2017 |
Drug-tolerant persister cancer cells in a high mesenchymal therapy-resistant state acquire dependency on GPX4 for survival; loss of GPX4 function causes selective ferroptotic death of persister cells in vitro and prevents tumor relapse in mice. |
GPX4 genetic knockdown/inhibition, ferroptosis cell death assays, in vivo mouse tumor relapse models |
Nature |
High |
29088702
|
| 2020 |
GPX4 deficiency enhances cellular lipid peroxidation, which leads to carbonylation of STING at C88 and inhibits STING trafficking from the ER to the Golgi complex, thereby specifically attenuating the cGAS-STING innate immune pathway; GPX4-maintained redox homeostasis is required for STING activation. |
GPX4 knockdown/knockout, carbonylation assays, subcellular fractionation and trafficking assays (ER-to-Golgi), viral infection models in vivo |
Nature immunology |
High |
32541831
|
| 2001 |
PHGPx (GPX4) acts as a protein thiol peroxidase in sperm, using its selenocysteine to oxidize specific capsule protein thiols via a peroxide-dependent mechanism, leading to cross-linking of mitochondrial capsule proteins and accounting for the selenium dependency of spermatogenesis. |
In vitro biochemical assays with H2O2, monobromobimane labeling, SDS-PAGE, mercaptoethanol release of active enzyme |
BioFactors (Oxford, England) |
High |
11568459
|
| 2021 |
GPX4 is the only enzyme capable of reducing esterified phospholipid hydroperoxides within a cellular context; a patient-derived R152H mutation destabilizes a critical loop adjacent to the active site, causing substantial loss of enzymatic function while paradoxically reducing GPX4 susceptibility to degradation. |
Structure-based analysis, cell models including patient fibroblasts, enzymatic activity assays, protein stability assays |
Nature chemical biology |
High |
34931062
|
| 2022 |
An allosteric site on GPX4 (distinct from the active site) is involved in native regeneration of GPX4 under low glutathione conditions; covalent binding of inhibitors to this site causes conformational change, inhibition of activity, and subsequent cellular GPX4 protein degradation. Co-crystal structures of six inhibitors bound in this site were determined. |
Co-crystal structure determination (X-ray), biophysical binding assays, enzymatic activity assays, compound library screen |
Cell chemical biology |
High |
36423641
|
| 2021 |
NMR characterization of GPX4 reveals an electrostatic (cationic patch) mechanism for membrane binding; mutagenesis identifies specific cationic patch residues required for membrane and cardiolipin headgroup binding, and a novel lipid binding site adjacent to the catalytic site. The cationic patch also mediates DNA binding, explaining the nuclear isoform's ability to target DNA-bound protamines. |
NMR spectroscopy (first NMR assignments of GPX4), site-directed mutagenesis, lipid/DNA binding assays |
Biochemistry |
High |
34492183
|
| 2003 |
Testis-specific nuclear form of GPX4 (PHGPx/snGPx) is generated by transcription initiation from an alternative promoter (not by alternative splicing) located in intron 1, producing an arginine-rich N-terminus responsible for nuclear localization and chromatin binding, with expression restricted to late stages of spermatogenesis. |
5'-RACE, RT-PCR, S1 nuclease analysis, in situ hybridization |
Biological chemistry |
High |
12751792
|
| 2023 |
Copper promotes GPX4 autophagic degradation and ferroptosis by directly binding to GPX4 cysteines C107 and C148, inducing GPX4 ubiquitination and aggregation; TAX1BP1 then acts as the autophagic receptor mediating GPX4 degradation. |
Co-immunoprecipitation, ubiquitination assays, autophagic flux assays, copper chelation experiments, mouse pancreatic cancer models |
Autophagy |
Medium |
36622894
|
| 2023 |
Creatine kinase B (CKB), when phosphorylated at T133 by AKT downstream of IGF1R signaling, acts as a non-canonical protein kinase to phosphorylate GPX4 at S104; this phosphorylation prevents HSC70 binding to GPX4 and blocks chaperone-mediated autophagy (CMA)-dependent GPX4 degradation, thereby suppressing ferroptosis. |
In vitro kinase assay, co-immunoprecipitation, CMA degradation assays, mutagenesis, mouse tumor models, human HCC specimen correlation |
Nature cell biology |
High |
37156912
|
| 2021 |
Legumain facilitates chaperone-mediated autophagy (CMA) of GPX4 in acute kidney injury: legumain directly interacts with HSC70, HSP90, and GPX4 (shown by immunoprecipitation), and legumain deficiency prevents CMA-dependent GPX4 degradation and tubular ferroptosis. |
Co-immunoprecipitation, lysosomal fractionation, proteasome/autophagy inhibitor experiments, legumain KO mice with IRI model |
Cell death & disease |
Medium |
33431801
|
| 2022 |
CST1 interacts with GPX4 (confirmed by Co-IP and mass spectrometry) and recruits the deubiquitinase OTUB1 to relieve GPX4 ubiquitination, thereby stabilizing GPX4 protein and inhibiting ferroptosis. |
Co-immunoprecipitation combined with mass spectrometry, ubiquitination assays, siRNA knockdown, in vivo mouse metastasis models |
Oncogene |
Medium |
36369321
|
| 2022 |
LUBAC (via its catalytic subunit HOIP) binds GPX4 and stabilizes it through linear (M1-linked) ubiquitination both under basal conditions and oxidative stress; LUBAC deficiency promotes GPX4 degradation and sensitizes cells to ferroptosis. |
Co-immunoprecipitation, linear ubiquitination assays, LUBAC genetic deficiency models, lipid peroxidation assays |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
36279464
|
| 2023 |
USP8 (ubiquitin-specific protease 8) directly interacts with and deubiquitinates GPX4, leading to GPX4 protein stabilization; USP8 inhibition destabilizes GPX4 and sensitizes cancer cells to ferroptosis. |
Co-immunoprecipitation, deubiquitination assays, intestinal epithelial cell-specific Usp8 knockout mice, in vivo tumor models |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
38598341
|
| 2023 |
OTUD5 acts as a deubiquitinase that binds GPX4, stabilizes it, and confers ferroptosis resistance; during ischemia-reperfusion, mTORC1-mediated autophagy degrades OTUD5, causing GPX4 decay and subsequent ferroptosis in renal tubular cells. |
Co-immunoprecipitation, ubiquitination assays, spatial transcriptomics, AAV-mediated OTUD5 delivery, mouse IRI model |
Nature communications |
Medium |
38110369
|
| 2023 |
TRIM26 directly interacts with GPX4 via its RING domain and catalyzes K63-linked ubiquitination of GPX4 at K107 and K117, switching polyubiquitination from K48 to K63 linkage and thereby enhancing GPX4 protein stability; PLK1-mediated S127 phosphorylation of TRIM26 enhances this interaction. |
Co-immunoprecipitation, ubiquitin linkage-specific assays (K48 vs K63), mutagenesis of ubiquitination sites, TRIM26 knockdown in glioma |
Cell death & disease |
Medium |
37872147
|
| 2025 |
GPX4 is S-palmitoylated on cysteine 66 by the acyltransferase ZDHHC20, which increases GPX4 protein stability; APT2 acts as the depalmitoylase of GPX4. Disrupting GPX4 palmitoylation sensitizes cancer cells to ferroptosis in vitro and in vivo. |
Palmitoylation assays (acyl-RAC), mutagenesis (C66), ZDHHC20/APT2 knockdown/inhibition, xenograft and orthotopic tumor models |
Nature communications |
High |
39833225
|
| 2025 |
ZDHHC8 palmitoylates GPX4 at Cys75, stabilizing GPX4 and suppressing ferroptosis; pharmacological inhibition of ZDHHC8 with PF-670462 promotes ZDHHC8 degradation, reduces GPX4 palmitoylation, and enhances ferroptosis sensitivity in tumor cells. |
Palmitoylation assays, mutagenesis (C75), small-molecule drug screening, xenograft tumor models, CD8+ T cell co-culture assays |
Nature cancer |
Medium |
40108413
|
| 2025 |
PRMT5 catalyzes symmetric dimethylation of GPX4 at arginine 152 (R152), which prevents Cullin1-FBW7 E3 ligase binding to GPX4 and blocks ubiquitination-mediated GPX4 degradation, thereby prolonging GPX4 half-life and suppressing ferroptosis. |
In vitro methylation assays, Co-immunoprecipitation, ubiquitination assays, PRMT5 inhibition, mutagenesis (R152), mouse tumor models |
Nature cell biology |
High |
40033101
|
| 2023 |
NEDD4L directly interacts with GPX4 (confirmed by Co-IP) and promotes GPX4 ubiquitination and degradation, facilitating granulosa cell ferroptosis and contributing to PCOS pathology. |
Co-immunoprecipitation, ubiquitination assays, CCK-8 cell viability, ferroptosis marker assays |
Endocrine connections |
Low |
36662677
|
| 2023 |
STUB1 is identified as an E3 ubiquitin ligase of GPX4, promoting GPX4 ubiquitination at site K191 and its degradation, thereby inducing ferroptosis in gastrointestinal stromal tumors. |
Co-immunoprecipitation, ubiquitination assays with site mutagenesis (K191), STUB1 knockdown and GPX4 overexpression, in vivo tumor models |
Cell death & disease |
Medium |
38110356
|
| 2024 |
OTUD5 deubiquitylates and stabilizes GPX4; p53 suppresses OTUD5 transcription, thereby promoting GPX4 ubiquitination and degradation and inducing ferroptosis in gastric cancer cells. |
Co-immunoprecipitation, deubiquitination assay, CRISPR-Cas9 Otud5 KO, luciferase reporter for p53-OTUD5 transcription, mouse tumor models |
Clinical and translational medicine |
Medium |
40070026
|
| 2026 |
FUNDC1 interacts with GPX4 via its 96–133 amino acid domain (shown by Co-IP), facilitating GPX4 recruitment from cytoplasm into mitochondria via the TOM/TIM import complex; within mitochondria, GPX4 is degraded by mitophagy, triggering hepatocyte ferroptosis. |
Co-immunoprecipitation (domain mapping), immunofluorescence colocalization, FUNDC1 knockout mouse (CCl4 model), mitochondrial fractionation |
Journal of advanced research |
Medium |
36828120
|
| 2023 |
STAT3 binds to the GPX4 promoter region and promotes its transcription (confirmed by ChIP and dual-luciferase reporter assay); thiostrepton blocks GPX4 expression by inhibiting STAT3, thereby inducing ferroptosis in pancreatic cancer cells. |
Chromatin immunoprecipitation (ChIP), dual-luciferase reporter assay, STAT3 inhibition, in vivo xenograft |
Cell death & disease |
Medium |
35859150
|
| 2024 |
Itaconate (produced by Irg1/aconitate decarboxylase 1) directly alkylates GPX4 at cysteine 66, allosterically enhancing GPX4 enzymatic activity, thereby protecting neurons from ferroptosis after intracerebral hemorrhage. |
Bioorthogonal itaconate-alkyne (ITalk) probe, GPX4 activity assay using phosphatidylcholine hydroperoxide, Irg1 transgenic/KO mouse models, in vitro cortical neuron ferroptosis assays |
Cell death and differentiation |
High |
38719928
|
| 1997 |
PHGPx (GPX4) shares an active site catalytic triad (selenocysteine, glutamine, tryptophan) with tetrameric GPXs; its unique ability to accommodate large lipophilic substrates (phospholipid hydroperoxides) is defined by structural and kinetic data. PHGPx also exhibits broad donor-substrate specificity (not restricted to GSH) and binds specific mitochondrial proteins via protein-protein interaction, suggesting a role in oxidizing specific protein thiols. |
Structural analysis, kinetic assays, protein-protein interaction (protein binding to mitochondrial proteins released by ionic strength and thiols) |
Biomedical and environmental sciences : BES |
Medium |
9315326
|
| 2003 |
Overexpression of mitochondrial PHGPx (mPHGPx) in neonatal rat cardiac myocytes reduces lipid peroxidation (malondialdehyde/HNE), decreases cytochrome c release from mitochondria, and preserves electron transport chain complex IV function following simulated ischemia/reoxygenation, demonstrating a direct mitochondrial protective role. |
Adenoviral overexpression of mPHGPx and cPHGPx, MDA/HNE quantification, cytochrome c release assay, complex IV activity assay, simulated I/R in neonatal cardiomyocytes |
Free radical biology & medicine |
Medium |
14583338
|
| 2021 |
GPX4 is required for follicular helper T (TFH) cell survival; selective deletion of GPX4 in T cells abrogates TFH cells and germinal center responses in immunized mice, establishing the selenium-GPX4-ferroptosis axis as a critical regulator of TFH homeostasis. |
T cell-specific GPX4 conditional knockout mice, immunization experiments, flow cytometry for TFH and germinal center B cells, selenium supplementation experiments |
Nature immunology |
High |
34413521
|
| 2022 |
GPX4 deficiency in macrophages and lung tissue results in ferroptotic cell death during Mycobacterium tuberculosis infection; Gpx4-deficient mice show increased lung necrosis and bacterial burden while Gpx4-overexpressing transgenic mice show decreased necrosis, establishing the GPX4/GSH axis as a host-resistance determinant in TB. |
Gpx4-deficient and Gpx4-transgenic mouse models, Mtb infection, ferrostatin-1 rescue in macrophages, lipid peroxidation assays |
The Journal of experimental medicine |
High |
36069923
|
| 2022 |
Adipocyte-specific GPX4 knockout spontaneously causes adipocyte hypertrophy, lipid peroxidation, inflammatory cytokine expression (TNF-α, IL-1β, IL-6, CXCL1), macrophage infiltration, glucose intolerance, and hepatic insulin resistance independent of ferroptosis-associated cell death. |
Conditional adipocyte-specific Gpx4 knockout (Adipoq-Cre and Fabp4-Cre), metabolic phenotyping (GTT, ITT), lipid peroxidation assays, cytokine measurement, TUNEL staining |
International journal of obesity (2005) |
High |
35031697
|
| 2024 |
PKA phosphorylates and promotes degradation of ALKBH5 (an m6A demethylase), thereby increasing m6A modification and stability of GPX4 mRNA; ALKBH5 deletion maintains GPX4 m6A modification and suppresses ferroptotic cell death, establishing a PKA-ALKBH5-GPX4 m6A regulatory axis. |
PKA kinase assays, ALKBH5 phosphorylation and degradation assays, m6A quantification of GPX4 mRNA, ALKBH5 knockout, ferroptosis cell death assays |
Cell death and differentiation |
Medium |
39901038
|
| 2023 |
RSL3 and ML162 (commonly used GPX4 inhibitors) completely lack capacity to inhibit recombinant selenoprotein GPX4 enzymatic activity in vitro; instead, they are efficient inhibitors of TXNRD1. This negative result suggests prior mechanistic studies attributing ferroptosis induction by these compounds solely to GPX4 inhibition need re-evaluation. |
In vitro enzymatic activity assays with recombinant GPX4 and TXNRD1, cell death assays with ferrostatin-1 rescue |
Redox biology |
Medium |
37087975
|
| 2025 |
TRAF6 ubiquitinates GPX4, promoting its recognition by the autophagic receptor p62 and selective autophagic degradation; a GPX4-targeted AUTAC exploiting this pathway induces GPX4 degradation and ferroptosis in breast cancer. |
Co-immunoprecipitation, ubiquitination assays, selective autophagy flux assays, AUTAC compound design, patient-derived organoids, in vivo tumor models |
Cell death and differentiation |
Medium |
40394165
|
| 2001 |
PHGPx (GPX4) was cloned as an anti-apoptotic and growth-promoting gene in Burkitt lymphoma cells, where cystine availability limiting glutathione biosynthesis determines cell susceptibility to oxidative stress-induced apoptosis, and PHGPx overexpression provides protection. |
Expression cloning in BL cells, overexpression rescue of low-density/low-serum-induced apoptosis |
BioFactors (Oxford, England) |
Medium |
11568455
|
| 1997 |
Co-transfection of both PHGPx cDNA and selenophosphate synthase (SelD) cDNA into endothelial cells produces significantly higher PHGPx activity than either alone, especially under selenium-limiting conditions, demonstrating that selenocysteine incorporation machinery (SelD) is rate-limiting for GPX4 expression and function. |
Stable transfection of endothelial cells (ECV304), PHGPx activity assays, hydroperoxide reduction assays, H2O2 cytotoxicity assays |
Biomedical and environmental sciences : BES |
Medium |
9315307
|