{"gene":"NOXA1","run_date":"2026-04-29T11:37:57","timeline":{"discoveries":[{"year":2007,"finding":"PKA phosphorylates NoxA1 at Ser172 and Ser461, promoting NoxA1 complex formation with 14-3-3 proteins and inducing dissociation of NoxA1 from the Nox1 complex at the plasma membrane, thereby inhibiting Nox1-dependent ROS production.","method":"Site-directed mutagenesis, phosphopeptide mapping, co-immunoprecipitation, cAMP manipulation in HEK293 and colon cell lines","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (mutagenesis, co-IP, functional ROS assay) in a single rigorous study","pmids":["17913709"],"is_preprint":false},{"year":2010,"finding":"MAP kinases phosphorylate NoxA1 at Ser282, and PKC and PKA phosphorylate Ser172; both phosphorylations decrease NoxA1 binding to NOX1 and Rac1, preventing NOX1 hyperactivation.","method":"In vitro kinase assay, site-directed mutagenesis, phosphopeptide mapping, ROS assay in HEK293 cells","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase assays combined with mutagenesis and functional ROS readout","pmids":["20110267"],"is_preprint":false},{"year":2010,"finding":"c-Src phosphorylates NoxA1 at Tyr110, and this phosphorylation is required for NoxA1 interaction with Tks4 and for Nox1-dependent ROS generation and functional invadopodia formation in human colon cancer cells.","method":"Phosphomimetic/unphosphorylable mutants, co-immunoprecipitation, ROS assay, ECM degradation assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1-2 — mutagenesis with multiple functional readouts (binding, ROS, invadopodia, ECM degradation)","pmids":["20943948"],"is_preprint":false},{"year":2006,"finding":"NOXA1 (the p67phox homologue) localizes to the cytosol of vascular smooth muscle cells and, upon co-expression with NOXO1, is targeted to the membrane; antisense knockdown of NOXA1 attenuates agonist-induced ROS production in mouse VSMC.","method":"Western blot fractionation, immunohistochemistry, fluorescent fusion protein co-expression, antisense knockdown with chemiluminescence ROS assay","journal":"Free radical biology & medicine","confidence":"Medium","confidence_rationale":"Tier 2-3 — direct localization with functional consequence via knockdown, single lab","pmids":["16814099"],"is_preprint":false},{"year":2008,"finding":"NoxA1 acts as an inhibitory regulator of Duox1 in airway epithelial cells; calcium-dependent dissociation of NoxA1 from plasma membrane-bound Duox activates H2O2 production, and NoxA1 knockdown increases basal H2O2 generation.","method":"Reconstituted cell-free system, mutational analysis, Duox1 knockdown in mucociliary airway epithelium model","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — reconstitution plus mutagenesis plus knockdown with functional readout in physiologically relevant model","pmids":["18606821"],"is_preprint":false},{"year":2013,"finding":"A peptide mimicking the activation domain of NOXA1 (NoxA1ds) selectively inhibits Nox1-derived superoxide by disrupting the binding interaction between Nox1 and NOXA1, with no effect on Nox2, Nox4, Nox5, or xanthine oxidase.","method":"Cell-free reconstituted Nox1 system, cytochrome c reduction, FRET (Nox1-YFP/NOXA1-CFP), ELISA, FRAP, confocal microscopy","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (FRET, FRAP, ELISA, cell-free assay) demonstrating direct binding disruption","pmids":["24187133"],"is_preprint":false},{"year":2010,"finding":"NOXO1 and NOXA1 both interact with membrane-bound p22phox; the C-terminal tail of NOXO1 competes for p22phox binding to regulate NOX1 complex assembly, and the NOXO1-NOXA1 interaction differs significantly from the p47phox-p67phox interaction.","method":"Isothermal titration calorimetry, pulldown assays","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1 — quantitative biophysical characterization (ITC) of protein-protein interactions","pmids":["20454568"],"is_preprint":false},{"year":2007,"finding":"The NOXA1 SH3 domain is not required for NOX1 activation but modulates kinetics of active complex formation; an alternatively spliced NOXA1 variant lacking the activation domain (NOXA1inhib) acts as a transdominant inhibitor and abolishes SH3 domain binding to NOXO1 and p47phox.","method":"Transfected K562 cells stably expressing NOX1/NOXO1, deletion/point mutants, kinetic superoxide generation assays, co-immunoprecipitation","journal":"Free radical biology & medicine","confidence":"High","confidence_rationale":"Tier 1-2 — systematic deletion/mutant analysis with kinetic functional readouts","pmids":["17602954"],"is_preprint":false},{"year":2012,"finding":"NOXA1 can activate Nox2 in a reconstituted cell-free system but with lower efficiency than p67phox; it shows higher EC50 values, lower Vmax (one-third of p67phox/p47phox), reduced FAD affinity for the oxidase, and less stable active complex formation.","method":"Purified component in vitro reconstitution with cyt.b558, Rac(Q61L), and Noxo1; kinetic analysis","journal":"Archives of biochemistry and biophysics","confidence":"High","confidence_rationale":"Tier 1 — purified component reconstitution with quantitative kinetic parameters","pmids":["22244833"],"is_preprint":false},{"year":2013,"finding":"PKC-dependent phosphorylation of NOXO1 at Thr341 enables sufficient interaction of NOXO1 with NOXA1, which is required for PMA-elicited enhancement of Nox1-catalyzed superoxide production.","method":"In vitro phosphorylation, alanine substitution mutagenesis, pulldown assays, superoxide production assays","journal":"The FEBS journal","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase assay with mutagenesis and binding/functional readouts","pmids":["23957209"],"is_preprint":false},{"year":2014,"finding":"PKCβ1 phosphorylates Nox1 at Thr429 in response to TNFα, and this phosphorylation facilitates the association of Nox1 with the NoxA1 activation domain, enabling NADPH oxidase complex assembly, ROS production, and vascular smooth muscle cell migration.","method":"Mass spectrometry, pharmacological inhibition, siRNA knockdown, site-directed mutagenesis, isothermal titration calorimetry","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 1 — ITC for binding affinity combined with mutagenesis and functional cellular assays","pmids":["25228390"],"is_preprint":false},{"year":2018,"finding":"NOXA1-dependent NOX1 activity in vascular smooth muscle cells mediates TNFα-induced ROS generation, VSMC proliferation and migration, and KLF4-mediated phenotypic switching to macrophage-like cells during atherogenesis.","method":"Systemic and SMC-specific Noxa1 knockout mice, vascular injury model, atherosclerosis model (Apoe-/-, Ldlr-/-), ROS assay, immunofluorescence","journal":"Redox biology","confidence":"High","confidence_rationale":"Tier 2 — clean conditional KO with multiple defined phenotypic and molecular readouts","pmids":["30576919"],"is_preprint":false},{"year":2022,"finding":"NOXA1/NOX1 signaling in renal principal cells of the distal nephron mediates angiotensin II-dependent activation of the epithelial sodium channel (ENaC), regulating Na+ reabsorption and blood pressure.","method":"Principal cell-specific Noxa1 knockout mice, patch-clamp electrophysiology on isolated split-opened tubules, NOX1-specific inhibitor (ML171), losartan","journal":"American journal of physiology. Renal physiology","confidence":"High","confidence_rationale":"Tier 2 — cell-type-specific KO with direct electrophysiological readout and pharmacological validation","pmids":["36201326"],"is_preprint":false},{"year":2022,"finding":"NOXA1/NOX1-dependent ROS in renal tubular cells enhances ENaC expression and Na+ reabsorption in response to angiotensin II and aldosterone; male-specific regulation is attributed to stronger NOXA1/NOX1-dependent ROS signaling.","method":"Whole-body and SMC-specific Noxa1 KO mice, telemetric blood pressure, Na+ excretion measurement, siRNA knockdown in renal epithelial cells","journal":"Antioxidants & redox signaling","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with multiple physiological and molecular readouts replicated across models","pmids":["34714114"],"is_preprint":false},{"year":2008,"finding":"NOXA1 is required for ROS generation induced by oxidized LDL (via LOX-1) and angiotensin II in human vascular endothelial cells; siRNA knockdown of NOXA1 or p22phox potently reduces these ROS responses.","method":"siRNA knockdown, dihydroethidium ROS assay, RT-PCR","journal":"Endothelium","confidence":"Medium","confidence_rationale":"Tier 3 — siRNA knockdown with ROS readout, single lab, single method","pmids":["18568954"],"is_preprint":false},{"year":2025,"finding":"NOXA1 suppresses ferroptosis in colorectal cancer cells by maintaining SLC7A11 and GPX4 expression; knockdown of NOXA1 decreases SLC7A11 and GPX4, elevates cellular ROS, induces ferroptosis, and sensitizes cells to radiotherapy.","method":"NOXA1 knockdown, Western blot, ROS assay, GSVA pathway analysis in CRC cell lines","journal":"International journal of medical sciences","confidence":"Low","confidence_rationale":"Tier 3 — single lab, single KD approach without mechanistic pathway resolution","pmids":["40084254"],"is_preprint":false},{"year":2025,"finding":"ERVWE1 upregulates NOXA1 transcription via USF2, and NOXA1 promotes macroautophagy (increased LC3B II/I ratio, autophagosome formation, reduced SQSTM1) while suppressing micromitophagy (reduced PINK1, Parkin, PDHA1 expression) in the context of schizophrenia-related neuronal cells.","method":"Cellular overexpression/knockdown, Western blot, luciferase reporter assay, bioinformatics","journal":"Virologica Sinica","confidence":"Low","confidence_rationale":"Tier 3 — single lab, mechanistic pathway placement based on knockdown without direct biochemical reconstitution","pmids":["40419114"],"is_preprint":false}],"current_model":"NOXA1 is a cytosolic activator subunit of the NOX1 NADPH oxidase complex that binds NOXO1 and Nox1 through its activation domain and SH3 domain to drive superoxide production; its activity is positively regulated by c-Src-mediated phosphorylation at Tyr110 (enabling Tks4 interaction and invadopodia formation) and negatively regulated by PKA- and PKC/MAPK-mediated phosphorylation at Ser172, Ser282, and Ser461 (which reduce binding to Nox1 and Rac1 and promote 14-3-3-dependent dissociation from the plasma membrane complex); additionally, NOXA1 plays an inhibitory role toward Duox1 in airway epithelium and mediates angiotensin II-dependent ENaC activation and Na+ homeostasis in the kidney via NOX1-derived ROS signaling."},"narrative":{"teleology":[{"year":2006,"claim":"Establishing that NOXA1 is a cytosolic activator that translocates to the membrane with NOXO1 and is required for agonist-induced ROS in vascular smooth muscle cells answered where and how NOXA1 participates in NOX1 complex assembly.","evidence":"Western blot fractionation, fluorescent fusion protein imaging, and antisense knockdown with ROS assay in mouse VSMC","pmids":["16814099"],"confidence":"Medium","gaps":["Single-lab antisense approach without genetic confirmation","Stimulus-specific versus constitutive role not delineated","Stoichiometry of NOXA1 in the membrane complex undefined"]},{"year":2007,"claim":"Dissecting NOXA1 domain requirements revealed that the activation domain is essential for NOX1 catalysis while the SH3 domain tunes kinetics, and identified a dominant-negative splice variant (NOXA1inhib) that blocks complex assembly.","evidence":"Systematic deletion/point mutants in K562 cells stably expressing NOX1/NOXO1 with kinetic superoxide assays and co-immunoprecipitation","pmids":["17602954"],"confidence":"High","gaps":["Structural basis of activation domain–Nox1 interface unresolved","Physiological relevance of NOXA1inhib splice variant not tested in vivo"]},{"year":2007,"claim":"Identification of PKA-mediated phosphorylation at Ser172 and Ser461 as a negative regulatory mechanism—promoting 14-3-3 binding and membrane dissociation—established how cAMP signaling restrains NOX1 activity.","evidence":"Site-directed mutagenesis, phosphopeptide mapping, co-IP, and cAMP manipulation in HEK293 and colon cell lines","pmids":["17913709"],"confidence":"High","gaps":["In vivo confirmation of 14-3-3 sequestration in tissues not provided","Identity of phosphatase(s) reversing these modifications unknown"]},{"year":2008,"claim":"Demonstrating that NOXA1 inhibits Duox1-dependent H₂O₂ in airway epithelium expanded NOXA1's role beyond NOX1 activation to a dual-oxidase inhibitory function governed by calcium-dependent dissociation.","evidence":"Reconstituted cell-free system, mutational analysis, and Duox1 knockdown in mucociliary airway epithelial model","pmids":["18606821"],"confidence":"High","gaps":["Binding interface between NOXA1 and Duox1 uncharacterized","Whether this inhibitory role extends to Duox2 untested"]},{"year":2010,"claim":"Mapping MAPK-dependent Ser282 and PKC-dependent Ser172 phosphorylation as additional negative regulatory sites that reduce Nox1 and Rac1 binding established a multi-kinase brake preventing NOX1 hyperactivation.","evidence":"In vitro kinase assays, site-directed mutagenesis, phosphopeptide mapping, and ROS assays in HEK293 cells","pmids":["20110267"],"confidence":"High","gaps":["Combinatorial effects of simultaneous multi-site phosphorylation not resolved","In vivo validation lacking"]},{"year":2010,"claim":"c-Src phosphorylation at Tyr110 was shown to be required for NOXA1–Tks4 interaction, Nox1-dependent ROS, and invadopodia formation, linking NOXA1 to cancer cell invasion.","evidence":"Phosphomimetic/unphosphorylable mutants, co-IP, ROS assay, and ECM degradation assay in human colon cancer cells","pmids":["20943948"],"confidence":"High","gaps":["Whether Tyr110 phosphorylation and Ser172/282/461 phosphorylation are coordinated remains unknown","In vivo tumor invasion phenotype not tested"]},{"year":2010,"claim":"Quantitative biophysical characterization showed that NOXA1 and NOXO1 both interact with p22phox, with NOXO1's C-terminal tail competing for p22phox binding—establishing how complex assembly is regulated at the membrane anchor.","evidence":"Isothermal titration calorimetry and pulldown assays","pmids":["20454568"],"confidence":"High","gaps":["Ternary complex structure of NOXA1–NOXO1–p22phox not determined","Relative affinities under physiological membrane conditions unknown"]},{"year":2013,"claim":"Development of the NoxA1ds peptide inhibitor that selectively disrupts the Nox1–NOXA1 activation domain interaction confirmed that this interface is the obligate catalytic trigger and provided an isoform-specific pharmacological tool.","evidence":"Cell-free Nox1 reconstitution, FRET, FRAP, ELISA, and confocal microscopy","pmids":["24187133"],"confidence":"High","gaps":["In vivo pharmacokinetics and efficacy of NoxA1ds not characterized in this study","Atomic-level binding interface undefined"]},{"year":2013,"claim":"PKC-dependent phosphorylation of NOXO1 at Thr341 was shown to be required for sufficient NOXO1–NOXA1 interaction and PMA-stimulated superoxide, revealing bidirectional phospho-regulation of the organizer–activator pair.","evidence":"In vitro phosphorylation, alanine mutagenesis, pulldown and superoxide assays","pmids":["23957209"],"confidence":"High","gaps":["Whether NOXO1 Thr341 and NOXA1 Ser172 phosphorylation are antagonistic in the same cell context unknown"]},{"year":2014,"claim":"PKCβ1-dependent Nox1 Thr429 phosphorylation was shown to enhance Nox1 association with the NOXA1 activation domain, demonstrating that signal-dependent modifications on Nox1 itself license activator recruitment.","evidence":"Mass spectrometry, ITC, siRNA, mutagenesis in TNFα-stimulated VSMC","pmids":["25228390"],"confidence":"High","gaps":["Whether other stimuli use the same Thr429 mechanism untested","Structural consequences of Thr429 phosphorylation unresolved"]},{"year":2018,"claim":"Conditional Noxa1 knockout in smooth muscle cells demonstrated that NOXA1/NOX1 drives TNFα-induced VSMC proliferation, migration, and KLF4-mediated macrophage-like phenotypic switching during atherogenesis in vivo.","evidence":"Systemic and SMC-specific Noxa1 KO mice in vascular injury and atherosclerosis (Apoe−/−, Ldlr−/−) models","pmids":["30576919"],"confidence":"High","gaps":["Whether endothelial NOXA1 contributes independently to atherogenesis not addressed","Downstream ROS targets mediating KLF4 induction unidentified"]},{"year":2022,"claim":"Cell-type-specific Noxa1 deletion and electrophysiology in kidney revealed that NOXA1/NOX1-derived ROS activate ENaC in distal nephron principal cells downstream of angiotensin II, establishing a renal sodium-handling function with sex-dependent regulation.","evidence":"Principal cell-specific Noxa1 KO mice, patch-clamp electrophysiology, NOX1 inhibitor ML171, losartan, telemetric blood pressure, Na+ excretion","pmids":["36201326","34714114"],"confidence":"High","gaps":["Direct ROS target linking NOX1 output to ENaC channel gating unidentified","Whether NOXA1 controls other renal ion channels unknown","Basis of sex-specific regulation at the molecular level unresolved"]},{"year":null,"claim":"A high-resolution structural model of the NOXA1–Nox1 activation domain interface is lacking, and the precise ROS-sensing intermediates downstream of NOXA1/NOX1 that trigger ENaC activation, KLF4 induction, and invadopodia signaling remain undefined.","evidence":"","pmids":[],"confidence":"High","gaps":["No crystal or cryo-EM structure of NOXA1 alone or in complex","Direct oxidation targets linking NOX1-derived superoxide to downstream effectors (ENaC, KLF4) uncharacterized","In vivo relevance of NOXA1 multi-site phosphorylation crosstalk not tested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,2,4,5,7]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,3]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,2,10,12,13]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[11]},{"term_id":"R-HSA-382551","term_label":"Transport of small molecules","supporting_discovery_ids":[12,13]}],"complexes":["NOX1 NADPH oxidase complex"],"partners":["NOX1","NOXO1","CYBA","RAC1","TKS4","YWHAZ","DUOX1"],"other_free_text":[]},"mechanistic_narrative":"NOXA1 is a cytosolic activator of the NOX1 NADPH oxidase complex that drives superoxide production in diverse cell types including vascular smooth muscle, colon epithelium, and renal tubular cells. NOXA1 binds Nox1 through its activation domain and associates with NOXO1 and p22phox to assemble the active membrane oxidase complex; its SH3 domain modulates the kinetics of complex formation rather than being strictly required for catalysis [PMID:17602954, PMID:20454568, PMID:24187133]. NOXA1 activity is positively regulated by c-Src phosphorylation at Tyr110 (enabling Tks4 binding and invadopodia formation) and negatively regulated by PKA/PKC/MAPK phosphorylation at Ser172, Ser282, and Ser461, which reduce Nox1 and Rac1 binding and promote 14-3-3-dependent sequestration away from the membrane [PMID:17913709, PMID:20110267, PMID:20943948]. Beyond NOX1 activation, NOXA1 inhibits Duox1-dependent H₂O₂ production in airway epithelium [PMID:18606821], mediates angiotensin II-dependent ENaC activation and sodium homeostasis in the distal nephron [PMID:36201326, PMID:34714114], and drives TNFα-induced VSMC phenotypic switching during atherogenesis [PMID:30576919]."},"prefetch_data":{"uniprot":{"accession":"Q86UR1","full_name":"NADPH oxidase activator 1","aliases":["Antigen NY-CO-31","NCF2-like protein","P67phox-like factor","p51-nox"],"length_aa":476,"mass_kda":50.9,"function":"Functions as an activator of NOX1, a superoxide-producing NADPH oxidase. Functions in the production of reactive oxygen species (ROS) which participate in a variety of biological processes including host defense, hormone biosynthesis, oxygen sensing and signal transduction. May also activate CYBB/gp91phox and NOX3","subcellular_location":"Cytoplasm; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q86UR1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NOXA1","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/NOXA1","total_profiled":1310},"omim":[{"mim_id":"611256","title":"NADPH OXIDASE ORGANIZER 1; NOXO1","url":"https://www.omim.org/entry/611256"},{"mim_id":"611255","title":"NADPH OXIDASE ACTIVATOR 1; NOXA1","url":"https://www.omim.org/entry/611255"},{"mim_id":"300951","title":"RING FINGER PROTEIN 113A; RNF113A","url":"https://www.omim.org/entry/300951"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NOXA1"},"hgnc":{"alias_symbol":["NY-CO-31","FLJ25475"],"prev_symbol":["SDCCAG31"]},"alphafold":{"accession":"Q86UR1","domains":[{"cath_id":"1.25.40.10","chopping":"5-170","consensus_level":"high","plddt":91.8895,"start":5,"end":170},{"cath_id":"3.10.20.90","chopping":"314-394","consensus_level":"medium","plddt":72.6526,"start":314,"end":394},{"cath_id":"2.30.30.40","chopping":"403-456","consensus_level":"medium","plddt":86.9607,"start":403,"end":456}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86UR1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86UR1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86UR1-F1-predicted_aligned_error_v6.png","plddt_mean":72.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NOXA1","jax_strain_url":"https://www.jax.org/strain/search?query=NOXA1"},"sequence":{"accession":"Q86UR1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86UR1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86UR1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86UR1"}},"corpus_meta":[{"pmid":"20943948","id":"PMC_20943948","title":"c-Src-mediated phosphorylation of NoxA1 and Tks4 induces the reactive oxygen species (ROS)-dependent formation of functional invadopodia in human colon cancer cells.","date":"2010","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/20943948","citation_count":91,"is_preprint":false},{"pmid":"17913709","id":"PMC_17913709","title":"Regulation of Nox1 activity via protein kinase A-mediated phosphorylation of NoxA1 and 14-3-3 binding.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17913709","citation_count":85,"is_preprint":false},{"pmid":"16814099","id":"PMC_16814099","title":"Noxa1 is a central component of the smooth muscle NADPH oxidase in mice.","date":"2006","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/16814099","citation_count":75,"is_preprint":false},{"pmid":"24187133","id":"PMC_24187133","title":"Selective recapitulation of conserved and nonconserved regions of putative NOXA1 protein activation domain confers isoform-specific inhibition of Nox1 oxidase and attenuation of endothelial cell migration.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24187133","citation_count":63,"is_preprint":false},{"pmid":"30576919","id":"PMC_30576919","title":"NOXA1-dependent NADPH oxidase regulates redox signaling and phenotype of vascular smooth muscle cell during atherogenesis.","date":"2018","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/30576919","citation_count":58,"is_preprint":false},{"pmid":"20110267","id":"PMC_20110267","title":"Phosphorylation of NADPH oxidase activator 1 (NOXA1) on serine 282 by MAP kinases and on serine 172 by protein kinase C and protein kinase A prevents NOX1 hyperactivation.","date":"2010","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/20110267","citation_count":52,"is_preprint":false},{"pmid":"20454568","id":"PMC_20454568","title":"Regulation of NOXO1 activity through reversible interactions with p22 and NOXA1.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/20454568","citation_count":51,"is_preprint":false},{"pmid":"18568954","id":"PMC_18568954","title":"Essential role of NOXA1 in generation of reactive oxygen species induced by oxidized low-density lipoprotein in human vascular endothelial cells.","date":"2008","source":"Endothelium : journal of endothelial cell research","url":"https://pubmed.ncbi.nlm.nih.gov/18568954","citation_count":39,"is_preprint":false},{"pmid":"18606821","id":"PMC_18606821","title":"Inhibitory action of NoxA1 on dual oxidase activity in airway cells.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18606821","citation_count":35,"is_preprint":false},{"pmid":"25228390","id":"PMC_25228390","title":"Phosphorylation of Nox1 regulates association with NoxA1 activation domain.","date":"2014","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/25228390","citation_count":30,"is_preprint":false},{"pmid":"23957209","id":"PMC_23957209","title":"Phosphorylation of Noxo1 at threonine 341 regulates its interaction with Noxa1 and the superoxide-producing activity of Nox1.","date":"2013","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/23957209","citation_count":28,"is_preprint":false},{"pmid":"34714114","id":"PMC_34714114","title":"Renal NOXA1/NOX1 Signaling Regulates Epithelial Sodium Channel and Sodium Retention in Angiotensin II-induced Hypertension.","date":"2022","source":"Antioxidants & redox signaling","url":"https://pubmed.ncbi.nlm.nih.gov/34714114","citation_count":25,"is_preprint":false},{"pmid":"17602954","id":"PMC_17602954","title":"NOX1 NADPH oxidase regulation by the NOXA1 SH3 domain.","date":"2007","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/17602954","citation_count":18,"is_preprint":false},{"pmid":"22244833","id":"PMC_22244833","title":"Noxa1 as a moderate activator of Nox2-based NADPH oxidase.","date":"2012","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/22244833","citation_count":16,"is_preprint":false},{"pmid":"36759130","id":"PMC_36759130","title":"Reactivity of renal and mesenteric resistance vessels to angiotensin II is mediated by NOXA1/NOX1 and superoxide signaling.","date":"2023","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36759130","citation_count":7,"is_preprint":false},{"pmid":"33708473","id":"PMC_33708473","title":"MiR-155 acts as an inhibitory factor in atherosclerosis-associated arterial pathogenesis by down-regulating NoxA1 related signaling pathway in ApoE-/- mouse.","date":"2021","source":"Cardiovascular diagnosis and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33708473","citation_count":7,"is_preprint":false},{"pmid":"36201326","id":"PMC_36201326","title":"NOXA1-dependent NADPH oxidase 1 signaling mediates angiotensin II activation of the epithelial sodium channel.","date":"2022","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36201326","citation_count":7,"is_preprint":false},{"pmid":"40419114","id":"PMC_40419114","title":"Human endogenous retrovirus W family envelope protein (ERVWE1) regulates macroautophagy activation and micromitophagy inhibition via NOXA1 in schizophrenia.","date":"2025","source":"Virologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/40419114","citation_count":2,"is_preprint":false},{"pmid":"40084254","id":"PMC_40084254","title":"NADPH oxidase activator 1 (NOXA1) suppresses ferroptosis and radiosensitization in colorectal cancer.","date":"2025","source":"International journal of medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40084254","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12976,"output_tokens":3806,"usd":0.048009},"stage2":{"model":"claude-opus-4-6","input_tokens":7249,"output_tokens":3132,"usd":0.171818},"total_usd":0.219827,"stage1_batch_id":"msgbatch_012ZiiCUBfB62NCTQd4Bk1Jo","stage2_batch_id":"msgbatch_01Xuqq9aUHoQouS7UVGZNStZ","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"PKA phosphorylates NoxA1 at Ser172 and Ser461, promoting NoxA1 complex formation with 14-3-3 proteins and inducing dissociation of NoxA1 from the Nox1 complex at the plasma membrane, thereby inhibiting Nox1-dependent ROS production.\",\n      \"method\": \"Site-directed mutagenesis, phosphopeptide mapping, co-immunoprecipitation, cAMP manipulation in HEK293 and colon cell lines\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (mutagenesis, co-IP, functional ROS assay) in a single rigorous study\",\n      \"pmids\": [\"17913709\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"MAP kinases phosphorylate NoxA1 at Ser282, and PKC and PKA phosphorylate Ser172; both phosphorylations decrease NoxA1 binding to NOX1 and Rac1, preventing NOX1 hyperactivation.\",\n      \"method\": \"In vitro kinase assay, site-directed mutagenesis, phosphopeptide mapping, ROS assay in HEK293 cells\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase assays combined with mutagenesis and functional ROS readout\",\n      \"pmids\": [\"20110267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"c-Src phosphorylates NoxA1 at Tyr110, and this phosphorylation is required for NoxA1 interaction with Tks4 and for Nox1-dependent ROS generation and functional invadopodia formation in human colon cancer cells.\",\n      \"method\": \"Phosphomimetic/unphosphorylable mutants, co-immunoprecipitation, ROS assay, ECM degradation assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis with multiple functional readouts (binding, ROS, invadopodia, ECM degradation)\",\n      \"pmids\": [\"20943948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"NOXA1 (the p67phox homologue) localizes to the cytosol of vascular smooth muscle cells and, upon co-expression with NOXO1, is targeted to the membrane; antisense knockdown of NOXA1 attenuates agonist-induced ROS production in mouse VSMC.\",\n      \"method\": \"Western blot fractionation, immunohistochemistry, fluorescent fusion protein co-expression, antisense knockdown with chemiluminescence ROS assay\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — direct localization with functional consequence via knockdown, single lab\",\n      \"pmids\": [\"16814099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"NoxA1 acts as an inhibitory regulator of Duox1 in airway epithelial cells; calcium-dependent dissociation of NoxA1 from plasma membrane-bound Duox activates H2O2 production, and NoxA1 knockdown increases basal H2O2 generation.\",\n      \"method\": \"Reconstituted cell-free system, mutational analysis, Duox1 knockdown in mucociliary airway epithelium model\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstitution plus mutagenesis plus knockdown with functional readout in physiologically relevant model\",\n      \"pmids\": [\"18606821\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"A peptide mimicking the activation domain of NOXA1 (NoxA1ds) selectively inhibits Nox1-derived superoxide by disrupting the binding interaction between Nox1 and NOXA1, with no effect on Nox2, Nox4, Nox5, or xanthine oxidase.\",\n      \"method\": \"Cell-free reconstituted Nox1 system, cytochrome c reduction, FRET (Nox1-YFP/NOXA1-CFP), ELISA, FRAP, confocal microscopy\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (FRET, FRAP, ELISA, cell-free assay) demonstrating direct binding disruption\",\n      \"pmids\": [\"24187133\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"NOXO1 and NOXA1 both interact with membrane-bound p22phox; the C-terminal tail of NOXO1 competes for p22phox binding to regulate NOX1 complex assembly, and the NOXO1-NOXA1 interaction differs significantly from the p47phox-p67phox interaction.\",\n      \"method\": \"Isothermal titration calorimetry, pulldown assays\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — quantitative biophysical characterization (ITC) of protein-protein interactions\",\n      \"pmids\": [\"20454568\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The NOXA1 SH3 domain is not required for NOX1 activation but modulates kinetics of active complex formation; an alternatively spliced NOXA1 variant lacking the activation domain (NOXA1inhib) acts as a transdominant inhibitor and abolishes SH3 domain binding to NOXO1 and p47phox.\",\n      \"method\": \"Transfected K562 cells stably expressing NOX1/NOXO1, deletion/point mutants, kinetic superoxide generation assays, co-immunoprecipitation\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — systematic deletion/mutant analysis with kinetic functional readouts\",\n      \"pmids\": [\"17602954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"NOXA1 can activate Nox2 in a reconstituted cell-free system but with lower efficiency than p67phox; it shows higher EC50 values, lower Vmax (one-third of p67phox/p47phox), reduced FAD affinity for the oxidase, and less stable active complex formation.\",\n      \"method\": \"Purified component in vitro reconstitution with cyt.b558, Rac(Q61L), and Noxo1; kinetic analysis\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — purified component reconstitution with quantitative kinetic parameters\",\n      \"pmids\": [\"22244833\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"PKC-dependent phosphorylation of NOXO1 at Thr341 enables sufficient interaction of NOXO1 with NOXA1, which is required for PMA-elicited enhancement of Nox1-catalyzed superoxide production.\",\n      \"method\": \"In vitro phosphorylation, alanine substitution mutagenesis, pulldown assays, superoxide production assays\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase assay with mutagenesis and binding/functional readouts\",\n      \"pmids\": [\"23957209\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"PKCβ1 phosphorylates Nox1 at Thr429 in response to TNFα, and this phosphorylation facilitates the association of Nox1 with the NoxA1 activation domain, enabling NADPH oxidase complex assembly, ROS production, and vascular smooth muscle cell migration.\",\n      \"method\": \"Mass spectrometry, pharmacological inhibition, siRNA knockdown, site-directed mutagenesis, isothermal titration calorimetry\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — ITC for binding affinity combined with mutagenesis and functional cellular assays\",\n      \"pmids\": [\"25228390\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NOXA1-dependent NOX1 activity in vascular smooth muscle cells mediates TNFα-induced ROS generation, VSMC proliferation and migration, and KLF4-mediated phenotypic switching to macrophage-like cells during atherogenesis.\",\n      \"method\": \"Systemic and SMC-specific Noxa1 knockout mice, vascular injury model, atherosclerosis model (Apoe-/-, Ldlr-/-), ROS assay, immunofluorescence\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO with multiple defined phenotypic and molecular readouts\",\n      \"pmids\": [\"30576919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NOXA1/NOX1 signaling in renal principal cells of the distal nephron mediates angiotensin II-dependent activation of the epithelial sodium channel (ENaC), regulating Na+ reabsorption and blood pressure.\",\n      \"method\": \"Principal cell-specific Noxa1 knockout mice, patch-clamp electrophysiology on isolated split-opened tubules, NOX1-specific inhibitor (ML171), losartan\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — cell-type-specific KO with direct electrophysiological readout and pharmacological validation\",\n      \"pmids\": [\"36201326\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NOXA1/NOX1-dependent ROS in renal tubular cells enhances ENaC expression and Na+ reabsorption in response to angiotensin II and aldosterone; male-specific regulation is attributed to stronger NOXA1/NOX1-dependent ROS signaling.\",\n      \"method\": \"Whole-body and SMC-specific Noxa1 KO mice, telemetric blood pressure, Na+ excretion measurement, siRNA knockdown in renal epithelial cells\",\n      \"journal\": \"Antioxidants & redox signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with multiple physiological and molecular readouts replicated across models\",\n      \"pmids\": [\"34714114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"NOXA1 is required for ROS generation induced by oxidized LDL (via LOX-1) and angiotensin II in human vascular endothelial cells; siRNA knockdown of NOXA1 or p22phox potently reduces these ROS responses.\",\n      \"method\": \"siRNA knockdown, dihydroethidium ROS assay, RT-PCR\",\n      \"journal\": \"Endothelium\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — siRNA knockdown with ROS readout, single lab, single method\",\n      \"pmids\": [\"18568954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NOXA1 suppresses ferroptosis in colorectal cancer cells by maintaining SLC7A11 and GPX4 expression; knockdown of NOXA1 decreases SLC7A11 and GPX4, elevates cellular ROS, induces ferroptosis, and sensitizes cells to radiotherapy.\",\n      \"method\": \"NOXA1 knockdown, Western blot, ROS assay, GSVA pathway analysis in CRC cell lines\",\n      \"journal\": \"International journal of medical sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single KD approach without mechanistic pathway resolution\",\n      \"pmids\": [\"40084254\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ERVWE1 upregulates NOXA1 transcription via USF2, and NOXA1 promotes macroautophagy (increased LC3B II/I ratio, autophagosome formation, reduced SQSTM1) while suppressing micromitophagy (reduced PINK1, Parkin, PDHA1 expression) in the context of schizophrenia-related neuronal cells.\",\n      \"method\": \"Cellular overexpression/knockdown, Western blot, luciferase reporter assay, bioinformatics\",\n      \"journal\": \"Virologica Sinica\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, mechanistic pathway placement based on knockdown without direct biochemical reconstitution\",\n      \"pmids\": [\"40419114\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NOXA1 is a cytosolic activator subunit of the NOX1 NADPH oxidase complex that binds NOXO1 and Nox1 through its activation domain and SH3 domain to drive superoxide production; its activity is positively regulated by c-Src-mediated phosphorylation at Tyr110 (enabling Tks4 interaction and invadopodia formation) and negatively regulated by PKA- and PKC/MAPK-mediated phosphorylation at Ser172, Ser282, and Ser461 (which reduce binding to Nox1 and Rac1 and promote 14-3-3-dependent dissociation from the plasma membrane complex); additionally, NOXA1 plays an inhibitory role toward Duox1 in airway epithelium and mediates angiotensin II-dependent ENaC activation and Na+ homeostasis in the kidney via NOX1-derived ROS signaling.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"NOXA1 is a cytosolic activator of the NOX1 NADPH oxidase complex that drives superoxide production in diverse cell types including vascular smooth muscle, colon epithelium, and renal tubular cells. NOXA1 binds Nox1 through its activation domain and associates with NOXO1 and p22phox to assemble the active membrane oxidase complex; its SH3 domain modulates the kinetics of complex formation rather than being strictly required for catalysis [PMID:17602954, PMID:20454568, PMID:24187133]. NOXA1 activity is positively regulated by c-Src phosphorylation at Tyr110 (enabling Tks4 binding and invadopodia formation) and negatively regulated by PKA/PKC/MAPK phosphorylation at Ser172, Ser282, and Ser461, which reduce Nox1 and Rac1 binding and promote 14-3-3-dependent sequestration away from the membrane [PMID:17913709, PMID:20110267, PMID:20943948]. Beyond NOX1 activation, NOXA1 inhibits Duox1-dependent H₂O₂ production in airway epithelium [PMID:18606821], mediates angiotensin II-dependent ENaC activation and sodium homeostasis in the distal nephron [PMID:36201326, PMID:34714114], and drives TNFα-induced VSMC phenotypic switching during atherogenesis [PMID:30576919].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Establishing that NOXA1 is a cytosolic activator that translocates to the membrane with NOXO1 and is required for agonist-induced ROS in vascular smooth muscle cells answered where and how NOXA1 participates in NOX1 complex assembly.\",\n      \"evidence\": \"Western blot fractionation, fluorescent fusion protein imaging, and antisense knockdown with ROS assay in mouse VSMC\",\n      \"pmids\": [\"16814099\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab antisense approach without genetic confirmation\", \"Stimulus-specific versus constitutive role not delineated\", \"Stoichiometry of NOXA1 in the membrane complex undefined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Dissecting NOXA1 domain requirements revealed that the activation domain is essential for NOX1 catalysis while the SH3 domain tunes kinetics, and identified a dominant-negative splice variant (NOXA1inhib) that blocks complex assembly.\",\n      \"evidence\": \"Systematic deletion/point mutants in K562 cells stably expressing NOX1/NOXO1 with kinetic superoxide assays and co-immunoprecipitation\",\n      \"pmids\": [\"17602954\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of activation domain–Nox1 interface unresolved\", \"Physiological relevance of NOXA1inhib splice variant not tested in vivo\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Identification of PKA-mediated phosphorylation at Ser172 and Ser461 as a negative regulatory mechanism—promoting 14-3-3 binding and membrane dissociation—established how cAMP signaling restrains NOX1 activity.\",\n      \"evidence\": \"Site-directed mutagenesis, phosphopeptide mapping, co-IP, and cAMP manipulation in HEK293 and colon cell lines\",\n      \"pmids\": [\"17913709\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo confirmation of 14-3-3 sequestration in tissues not provided\", \"Identity of phosphatase(s) reversing these modifications unknown\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Demonstrating that NOXA1 inhibits Duox1-dependent H₂O₂ in airway epithelium expanded NOXA1's role beyond NOX1 activation to a dual-oxidase inhibitory function governed by calcium-dependent dissociation.\",\n      \"evidence\": \"Reconstituted cell-free system, mutational analysis, and Duox1 knockdown in mucociliary airway epithelial model\",\n      \"pmids\": [\"18606821\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Binding interface between NOXA1 and Duox1 uncharacterized\", \"Whether this inhibitory role extends to Duox2 untested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Mapping MAPK-dependent Ser282 and PKC-dependent Ser172 phosphorylation as additional negative regulatory sites that reduce Nox1 and Rac1 binding established a multi-kinase brake preventing NOX1 hyperactivation.\",\n      \"evidence\": \"In vitro kinase assays, site-directed mutagenesis, phosphopeptide mapping, and ROS assays in HEK293 cells\",\n      \"pmids\": [\"20110267\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Combinatorial effects of simultaneous multi-site phosphorylation not resolved\", \"In vivo validation lacking\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"c-Src phosphorylation at Tyr110 was shown to be required for NOXA1–Tks4 interaction, Nox1-dependent ROS, and invadopodia formation, linking NOXA1 to cancer cell invasion.\",\n      \"evidence\": \"Phosphomimetic/unphosphorylable mutants, co-IP, ROS assay, and ECM degradation assay in human colon cancer cells\",\n      \"pmids\": [\"20943948\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Tyr110 phosphorylation and Ser172/282/461 phosphorylation are coordinated remains unknown\", \"In vivo tumor invasion phenotype not tested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Quantitative biophysical characterization showed that NOXA1 and NOXO1 both interact with p22phox, with NOXO1's C-terminal tail competing for p22phox binding—establishing how complex assembly is regulated at the membrane anchor.\",\n      \"evidence\": \"Isothermal titration calorimetry and pulldown assays\",\n      \"pmids\": [\"20454568\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ternary complex structure of NOXA1–NOXO1–p22phox not determined\", \"Relative affinities under physiological membrane conditions unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Development of the NoxA1ds peptide inhibitor that selectively disrupts the Nox1–NOXA1 activation domain interaction confirmed that this interface is the obligate catalytic trigger and provided an isoform-specific pharmacological tool.\",\n      \"evidence\": \"Cell-free Nox1 reconstitution, FRET, FRAP, ELISA, and confocal microscopy\",\n      \"pmids\": [\"24187133\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo pharmacokinetics and efficacy of NoxA1ds not characterized in this study\", \"Atomic-level binding interface undefined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"PKC-dependent phosphorylation of NOXO1 at Thr341 was shown to be required for sufficient NOXO1–NOXA1 interaction and PMA-stimulated superoxide, revealing bidirectional phospho-regulation of the organizer–activator pair.\",\n      \"evidence\": \"In vitro phosphorylation, alanine mutagenesis, pulldown and superoxide assays\",\n      \"pmids\": [\"23957209\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NOXO1 Thr341 and NOXA1 Ser172 phosphorylation are antagonistic in the same cell context unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"PKCβ1-dependent Nox1 Thr429 phosphorylation was shown to enhance Nox1 association with the NOXA1 activation domain, demonstrating that signal-dependent modifications on Nox1 itself license activator recruitment.\",\n      \"evidence\": \"Mass spectrometry, ITC, siRNA, mutagenesis in TNFα-stimulated VSMC\",\n      \"pmids\": [\"25228390\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other stimuli use the same Thr429 mechanism untested\", \"Structural consequences of Thr429 phosphorylation unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Conditional Noxa1 knockout in smooth muscle cells demonstrated that NOXA1/NOX1 drives TNFα-induced VSMC proliferation, migration, and KLF4-mediated macrophage-like phenotypic switching during atherogenesis in vivo.\",\n      \"evidence\": \"Systemic and SMC-specific Noxa1 KO mice in vascular injury and atherosclerosis (Apoe−/−, Ldlr−/−) models\",\n      \"pmids\": [\"30576919\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether endothelial NOXA1 contributes independently to atherogenesis not addressed\", \"Downstream ROS targets mediating KLF4 induction unidentified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Cell-type-specific Noxa1 deletion and electrophysiology in kidney revealed that NOXA1/NOX1-derived ROS activate ENaC in distal nephron principal cells downstream of angiotensin II, establishing a renal sodium-handling function with sex-dependent regulation.\",\n      \"evidence\": \"Principal cell-specific Noxa1 KO mice, patch-clamp electrophysiology, NOX1 inhibitor ML171, losartan, telemetric blood pressure, Na+ excretion\",\n      \"pmids\": [\"36201326\", \"34714114\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct ROS target linking NOX1 output to ENaC channel gating unidentified\", \"Whether NOXA1 controls other renal ion channels unknown\", \"Basis of sex-specific regulation at the molecular level unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A high-resolution structural model of the NOXA1–Nox1 activation domain interface is lacking, and the precise ROS-sensing intermediates downstream of NOXA1/NOX1 that trigger ENaC activation, KLF4 induction, and invadopodia signaling remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No crystal or cryo-EM structure of NOXA1 alone or in complex\", \"Direct oxidation targets linking NOX1-derived superoxide to downstream effectors (ENaC, KLF4) uncharacterized\", \"In vivo relevance of NOXA1 multi-site phosphorylation crosstalk not tested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 2, 4, 5, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 10, 12, 13]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [11]},\n      {\"term_id\": \"R-HSA-382551\", \"supporting_discovery_ids\": [12, 13]}\n    ],\n    \"complexes\": [\n      \"NOX1 NADPH oxidase complex\"\n    ],\n    \"partners\": [\n      \"NOX1\",\n      \"NOXO1\",\n      \"CYBA\",\n      \"RAC1\",\n      \"TKS4\",\n      \"YWHAZ\",\n      \"DUOX1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}