Affinage

FDX2

Ferredoxin-2, mitochondrial · UniProt Q6P4F2

Length
183 aa
Mass
19.5 kDa
Annotated
2026-06-09
18 papers in source corpus 11 papers cited in narrative 11 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

FDX2 is a mitochondrial [2Fe-2S] ferredoxin that serves as the dedicated electron donor for iron-sulfur (Fe-S) cluster biogenesis, distinct from its paralog FDX1, which instead supports steroidogenesis, heme a biosynthesis, and lipoyl cofactor synthesis (PMID:20547883, PMID:36280795). FDX2 acts at two sequential steps: it donates electrons to the NFS1/ISD11/Acp cysteine desulfurase complex—binding near its Fe-S cluster more tightly than FDX1 and driving conversion of L-cysteine to L-alanine and sulfide to assemble [2Fe-2S] clusters on ISCU (PMID:28001042), and it subsequently provides electrons for reductive [2Fe-2S] fusion on ISCA1-ISCA2 in an IBA57-dependent manner to mature [4Fe-4S] proteins, a step for which no other cellular reductant substitutes (PMID:32817474). FDX2 receives electrons from the NADPH-coupled reductase FDXR, and its C-terminal tail mediates this electron transfer; substrate specificity that separates FDX2 from FDX1 is encoded by small conserved sequence motifs, since swapping them exchanges target specificity (PMID:36280795, PMID:39467201). By competing with frataxin for the NFS1 binding site, FDX2 levels tune cysteine desulfurase output, and reducing FDX2 dosage can ameliorate frataxin-deficiency phenotypes (PMID:41372402). Loss of FDX2 collapses Fe-S protein maturation, causes mitochondrial iron accumulation, and triggers p53-dependent senescence, apoptosis, and ferroptosis sensitization (PMID:20547883, PMID:39151727). Loss-of-function and missense FDX2 mutations cause a human mitochondrial disease marked by Fe-S-dependent respiratory chain and aconitase deficiencies, muscle iron accumulation, and impaired mitochondrial respiration (PMID:24281368, PMID:30010796, PMID:41372147).

Mechanistic history

Synthesis pass · year-by-year structured walk · 11 steps
  1. 2010 High

    Established that the two human mitochondrial ferredoxins are functionally non-redundant, assigning FDX2 specifically to Fe-S protein and heme A biogenesis rather than steroidogenesis.

    Evidence RNAi depletion of FDX1 vs FDX2 in human cells with biochemical readouts for Fe-S assembly, heme A, steroid conversion, and iron homeostasis

    PMID:20547883

    Open questions at the time
    • Did not resolve which biosynthetic steps FDX2 acts on
    • No structural basis for substrate discrimination
  2. 2013 Medium

    Linked FDX2 to human disease by showing a loss-of-function initiation-codon mutation depletes the protein and cripples Fe-S-dependent respiratory complexes, validating the in vivo requirement for FDX2 in Fe-S biogenesis.

    Evidence Exome sequencing/homozygosity mapping plus enzyme activity assays and western blot in patient muscle and fibroblasts

    PMID:24281368

    Open questions at the time
    • Single family/lab
    • No reconstitution of the mechanistic defect
    • Genotype-phenotype spectrum unknown
  3. 2017 High

    Defined the first molecular step of FDX2 action, showing it binds the cysteine desulfurase complex near its Fe-S cluster and donates electrons to assemble [2Fe-2S] clusters on ISCU faster than FDX1.

    Evidence NMR interaction mapping, isothermal titration calorimetry, and in vitro Fe-S assembly assays on ISCU

    PMID:28001042

    Open questions at the time
    • In vitro context only
    • Did not address downstream [4Fe-4S] maturation
    • Physiological reductant FDXR not yet incorporated
  4. 2018 Medium

    Identified the recurrent P144L missense allele as disease-causing, reinforcing FDX2's essential in vivo role and implicating a specific residue in protein function.

    Evidence Whole exome sequencing, RT-PCR, western blot, and histochemical/iron staining of patient muscle

    PMID:30010796

    Open questions at the time
    • Mechanism by which P144L reduces protein/function not defined
    • Single lab
  5. 2020 High

    Revealed a second, distinct electron-transfer role for FDX2 in reductive [2Fe-2S] fusion on ISCA1-ISCA2 to build [4Fe-4S] clusters, showing FDX1 and other reductants cannot substitute.

    Evidence In vitro reconstitution of [4Fe-4S] aconitase maturation with defined components, Mössbauer spectroscopy, and complementation assays

    PMID:32817474

    Open questions at the time
    • Structural basis of ISCA1-ISCA2/IBA57 recognition unresolved
    • Coupling between the two FDX2 steps in cells not defined
  6. 2022 High

    Pinpointed that small conserved sequence motifs encode the substrate specificity dividing FDX2 (Fe-S maturation) from FDX1 (steroidogenesis, heme a, lipoylation), demonstrated by motif swapping.

    Evidence RNAi depletion, in vitro assays, and domain-swap mutagenesis with functional readouts for each pathway

    PMID:36280795

    Open questions at the time
    • Atomic-level mechanism of motif-encoded recognition not resolved
    • How motifs dictate partner binding unknown
  7. 2024 High

    Provided the mechanistic explanation for the P144L disease allele, showing it disrupts FDX2-FDXR recognition and electron transfer, thereby impairing both [2Fe-2S] and [4Fe-4S] assembly, and assigned a functional role to the FDX2 C-terminal tail.

    Evidence NMR/EPR structural characterization, redox potentiometry, and in vitro electron transfer assays comparing WT vs P144L FDX2 with FDXR

    PMID:39467201

    Open questions at the time
    • In cellulo confirmation of the FDXR-interface defect limited
    • Full FDX2-FDXR complex structure not determined
  8. 2024 Medium

    Connected FDX2 loss to cell-fate decisions, showing collapse of Fe-S proteins and iron overload triggers p53-dependent senescence (or apoptosis when p53 is absent) and sensitizes cells to ferroptosis.

    Evidence Conditional FDX2 knockout in ovarian cancer cells with proteomics, DNA damage assays, and ferroptosis/p53 pathway analysis

    PMID:39151727

    Open questions at the time
    • Single cancer cell context
    • Direct link between specific Fe-S client loss and senescence not dissected
  9. 2025 High

    Established that FDX2 and frataxin compete for the same NFS1 binding site, identifying FDX2 dosage as a therapeutic lever in Friedreich's ataxia.

    Evidence Forward genetic screen in C. elegans, in vitro NFS1 activity assays with excess FDX2, mammalian Fe-S synthesis assays, and mouse Friedreich's ataxia rescue

    PMID:41372402

    Open questions at the time
    • Structural detail of the shared NFS1 interface not fully resolved
    • Therapeutic window of FDX2 reduction in humans untested
  10. 2025 Medium

    Characterized the electronic structure of the FDX2 [Fe2S2]2+ cluster, showing the two Fe(III) centers are inequivalent due to spin density transfer via C-H---S-Fe interactions.

    Evidence Paramagnetic NMR combined with DFT quantum chemical calculations

    PMID:40121555

    Open questions at the time
    • No functional mutagenesis validation
    • Relevance of cluster electronics to electron-transfer kinetics not tested
  11. 2025 Medium

    Extended the disease spectrum with a splicing mutation that alters N-terminal residues while preserving fold, linking low FDX2 levels to impaired respiration, Fe-S defects, iron accumulation, and reduced SOD2.

    Evidence RNA splicing analysis, NMR structural comparison, mitochondrial respiration and Fe-S activity assays, and SOD2 western blot in patient cells

    PMID:41372147

    Open questions at the time
    • Single patient
    • Mechanism of SOD2 reduction not established

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the two spatially and temporally distinct FDX2 electron-donation steps are coordinated in vivo, and the high-resolution structure of FDX2 in complex with FDXR and with the NFS1 desulfurase machinery, remain undefined.
  • No full structure of FDX2-FDXR or FDX2-NFS1 complexes
  • Regulation of FDX2 partitioning between ISCU and ISCA steps unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016491 oxidoreductase activity 3 GO:0140104 molecular carrier activity 2
Localization
GO:0005739 mitochondrion 4
Pathway
R-HSA-1430728 Metabolism 3 R-HSA-5357801 Programmed Cell Death 1

Evidence

Reading pass · 11 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2010 FDX2 (Fdx2), but not FDX1 (Fdx1/adrenodoxin), is essential for heme A and Fe/S protein biosynthesis in human mitochondria. FDX2 deficiency impairs Fe/S protein biogenesis, leading to increased cellular iron uptake and iron accumulation in mitochondria. Conversely, FDX2 is unable to efficiently reduce mitochondrial cytochromes P450 or convert steroids, functions specific to FDX1. RNAi-mediated depletion of FDX1 or FDX2 in human cells, with biochemical readouts for heme A, Fe/S cluster assembly, steroid conversion, and iron homeostasis Proceedings of the National Academy of Sciences of the United States of America High 20547883
2017 Both FDX1 and FDX2 bind the cysteine desulfurase complex (NFS1/ISD11/Acp) via residues near their Fe-S clusters, but FDX2 binds the complex more tightly than FDX1 (by isothermal titration calorimetry). In vitro, reduced FDX2 supports Fe-S cluster assembly on ISCU at a faster rate than FDX1. FDX2 donates electrons to the cysteine desulfurase complex, resulting in conversion of L-cysteine to L-alanine and sulfide generation. NMR spectroscopy (protein-protein interaction mapping), isothermal titration calorimetry, in vitro Fe-S cluster assembly assay on ISCU Biochemistry High 28001042
2020 FDX2, together with its reductase FDXR (NADPH-coupled), provides electrons required for reductive [2Fe-2S] cluster fusion on ISCA1-ISCA2 in an IBA57-dependent manner, enabling maturation of mitochondrial [4Fe-4S] proteins. This electron transfer step is distinct from FDX2's earlier role in [2Fe-2S] cluster synthesis on ISCU2. FDX1 and other cellular reducing systems cannot substitute for FDX2 in this step. In vitro reconstitution of [4Fe-4S] aconitase maturation without artificial reductants, using purified components; Mössbauer spectroscopy; biochemical complementation assays Proceedings of the National Academy of Sciences of the United States of America High 32817474
2022 FDX2, but not FDX1, is required for Fe-S protein maturation; FDX1 is specific for steroidogenesis, heme a biosynthesis, and lipoyl cofactor biosynthesis (providing electrons to lipoyl synthase). The distinct substrate specificity of each ferredoxin is determined by small conserved sequence motifs; swapping these motifs exchanges their target specificities. RNAi depletion in human cells, in vitro biochemical assays, domain-swap mutagenesis, functional assays for steroidogenesis/lipoylation/Fe-S assembly Nature chemical biology High 36280795
2013 A homozygous loss-of-function mutation in FDX1L (FDX2; c.1A>T disrupting the ATG initiation codon) causes severe reduction of Fdx2 protein in patient muscle and fibroblast mitochondria, resulting in severely impaired activities of Fe-S-dependent respiratory chain complexes I, II, III and mitochondrial aconitase, establishing FDX2 as the second component of the Fe-S cluster biogenesis machinery in human muscle. Exome sequencing + homozygosity mapping; enzyme activity assays in patient skeletal muscle; western blot of patient mitochondria European journal of human genetics : EJHG Medium 24281368
2018 A homozygous missense mutation in FDX2 (c.431C>T, p.P144L) causes severely reduced FDX2 protein expression (normal mRNA, reduced protein by western blot) in patient muscle, with an unusual pattern of succinate dehydrogenase and cytochrome c oxidase deficiency and iron accumulation on muscle biopsy, confirming FDX2's essential role in Fe-S cluster biogenesis in vivo. Genetic mapping + whole exome sequencing; RT-PCR; western blot of patient muscle; muscle biopsy with histochemical and iron staining Brain : a journal of neurology Medium 30010796
2024 The pathogenic P144L mutation of FDX2 negatively affects the FDXR-dependent electron transfer pathway from NADPH to FDX2 by altering the protein-protein recognition between FDX2 and its physiological electron donor FDXR, thereby reducing FDX2's capacity to assemble both [2Fe-2S] and [4Fe-4S] clusters. The C-terminal tail of FDX2 plays a functional role in electron transfer between FDX2 and FDXR. Structural characterization (NMR, EPR), redox potentiometry, in vitro electron transfer assays comparing WT and P144L FDX2 with FDXR, protein-protein interaction mapping Protein science : a publication of the Protein Society High 39467201
2025 Dominant gain-of-function mutations at the FDX2-NFS1 binding interface (identified in C. elegans and validated biochemically) suppress frataxin deficiency by boosting iron-sulfur cluster levels. Excess wild-type FDX2 inhibits frataxin-stimulated NFS1 (cysteine desulfurase) activity in vitro and blocks Fe-S cluster synthesis in mammalian cell culture, indicating that frataxin and FDX2 compete for the same binding site on NFS1. Partial knockdown of FDX2 (loss of one gene copy) ameliorates the growth defect in frataxin-mutant C. elegans and the ataxia phenotype in a mouse model of Friedreich's ataxia. Genome-scale forward genetic screen in C. elegans; in vitro NFS1 activity assays with excess FDX2; mammalian cell culture Fe-S cluster synthesis assay; mouse model rescue experiment Nature High 41372402
2024 FDX2 loss in ovarian cancer cells causes global downregulation of Fe-S-containing proteins and Fe2+ overload, resulting in DNA damage and p53 pathway activation, driving senescence. In p53-deficient cells, FDX2 loss leads to apoptosis rather than senescence. FDX2 loss also sensitizes cells to ferroptosis via compromised redox homeostasis of membrane phospholipids. Conditional knockout of FDX2 in ovarian cancer cell line; proteomics; DNA damage assays; ferroptosis sensitivity assays; p53 pathway analysis The Journal of biological chemistry Medium 39151727
2025 Paramagnetic NMR combined with DFT calculations of the [Fe2S2]2+ cluster of human FDX2 reveals that the two Fe(III) centers are inequivalent due to electron spin density transfer between cluster inorganic sulfide ions and aliphatic carbon atoms via C-H---S-Fe3+ interactions. The magnetic exchange coupling constant between the two Fe3+ ions is estimated at ~386 cm-1. Paramagnetic NMR spectroscopy; density functional theory quantum chemical calculations Inorganic chemistry Medium 40121555
2025 A novel FDX2 splicing mutation (c.200+4 A>G) generates a mutant protein with 21 replacement N-terminal residues (replacing exon-2-encoded residues) that likely retains structural integrity (no significant backbone dynamic differences vs WT by NMR), but patient cells with low FDX2 levels show impaired mitochondrial respiration, defects in Fe-S proteins, enhanced mitochondrial iron accumulation, and significantly diminished mitochondrial SOD2 levels. RNA splicing analysis; NMR structural comparison of mutant vs WT FDX2; mitochondrial respiration assay; Fe-S protein activity assays; mitochondrial iron measurement; western blot for SOD2 Cell death & disease Medium 41372147

Source papers

Stage 0 corpus · 18 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2010 Humans possess two mitochondrial ferredoxins, Fdx1 and Fdx2, with distinct roles in steroidogenesis, heme, and Fe/S cluster biosynthesis. Proceedings of the National Academy of Sciences of the United States of America 321 20547883
2022 Functional spectrum and specificity of mitochondrial ferredoxins FDX1 and FDX2. Nature chemical biology 130 36280795
2017 Human Mitochondrial Ferredoxin 1 (FDX1) and Ferredoxin 2 (FDX2) Both Bind Cysteine Desulfurase and Donate Electrons for Iron-Sulfur Cluster Biosynthesis. Biochemistry 117 28001042
2020 Mitochondrial [4Fe-4S] protein assembly involves reductive [2Fe-2S] cluster fusion on ISCA1-ISCA2 by electron flow from ferredoxin FDX2. Proceedings of the National Academy of Sciences of the United States of America 71 32817474
2013 Deleterious mutation in FDX1L gene is associated with a novel mitochondrial muscle myopathy. European journal of human genetics : EJHG 70 24281368
2018 A novel complex neurological phenotype due to a homozygous mutation in FDX2. Brain : a journal of neurology 41 30010796
2015 Crystal structure and biochemical characterization of Chlamydomonas FDX2 reveal two residues that, when mutated, partially confer FDX2 the redox potential and catalytic properties of FDX1. Photosynthesis research 17 26526668
2022 FDX2 and ISCU Gene Variations Lead to Rhabdomyolysis With Distinct Severity and Iron Regulation. Neurology. Genetics 15 35079622
2024 FDX2, an iron-sulfur cluster assembly factor, is essential to prevent cellular senescence, apoptosis or ferroptosis of ovarian cancer cells. The Journal of biological chemistry 12 39151727
2024 Unraveling the molecular determinants of a rare human mitochondrial disorder caused by the P144L mutation of FDX2. Protein science : a publication of the Protein Society 8 39467201
2021 Rare presentation of FDX2-related disorder and untargeted global metabolomics findings. American journal of medical genetics. Part A 7 34905296
2024 Clinical, biochemical and molecular characterization of a new case with FDX2-related mitochondrial disorder: Potential biomarkers and treatment options. JIMD reports 6 38444577
2023 Α rare case of myopathy, lactic acidosis, and severe rhabdomyolysis, due to a homozygous mutation of the ferredoxin-2 (FDX2) gene. American journal of medical genetics. Part A 5 37565517
2025 Shedding Light on the Electron Delocalization Pathway at the [Fe2S2]2+ Cluster of FDX2. Inorganic chemistry 3 40121555
2025 Mutations in mitochondrial ferredoxin FDX2 suppress frataxin deficiency. Nature 3 41372402
2026 Client distribution between Chlamydomonas FDX1 and FDX2 in carbon, nitrogen and sulfur assimilation. bioRxiv : the preprint server for biology 1 41648530
2026 The iron-sulfur cluster assembly factor FDX2 is required for tumor initiation but not for growth of established tumors in transplantation models. The Journal of biological chemistry 0 42208897
2025 A novel mutation in FDX2 provides insights into the pathogenesis of MEOAL mitochondrial neuromuscular disease. Cell death & disease 0 41372147

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