Affinage

Showing NCF2 — P67PHOX is a alias.

NCF2

Neutrophil cytosol factor 2 · UniProt P19878

Length
526 aa
Mass
59.8 kDa
Annotated
2026-06-10
100 papers in source corpus 42 papers cited in narrative 42 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NCF2 (p67phox) is an essential cytosolic activator of the phagocyte NADPH oxidase, functioning as a multidomain scaffold that both directly activates the catalytic flavocytochrome b558 and bridges the other cytosolic oxidase factors during assembly (PMID:19723583, PMID:11796733, PMID:7896790). Its N-terminal TPR domain binds GTP-loaded Rac (preferentially Rac2 over Rac1) through a novel TPR-insertion interaction mode, establishing p67phox as the Rac effector in the oxidase (PMID:8036496, PMID:11090627, PMID:8550629); Rac plays a dual role, tethering p67phox to the membrane and inducing an activating conformational change in the activation domain (PMID:14761978). A discrete activation domain spanning residues 199–210 (critical residue V204) directly stimulates the reductive half-reaction of FAD—hydride transfer from NADPH to flavin—with p67phox alone sufficing for FAD reduction while p47phox is required for downstream electron transfer to heme (PMID:9642219, PMID:10438466, PMID:7896790). In the resting cytosol p67phox is the central organizer of a 1:1:1 heterotrimer, binding p47phox and bridging p40phox via a PC-motif interaction with the region between its two SH3 domains (PMID:11796733, PMID:9490029). Oxidase activation requires phosphorylation—principally at Thr233—by PKC isoforms (PKCδ in monocytes, PKCβ elsewhere), ERK2, and p38MAPK, together with membrane translocation that depends on p40phox or p47phox as carrier proteins delivering p67phox to early endosomes or the plasma membrane respectively, and on intact cytochrome b558 for stable membrane retention (PMID:9931304, PMID:17122360, PMID:7964505, PMID:10233905, PMID:15591124). NCF2 transcription is controlled by a PU.1/IRF1/ICSBP/CBP module activated by IFN-γ, repressed by HoxA10, and induced by p53 (PMID:10570299, PMID:16210632, PMID:23187810). Loss-of-function NCF2 mutations cause chronic granulomatous disease, and a rare variant reducing RAC2 binding is associated with very early onset inflammatory bowel disease (PMID:8879195, PMID:21900546).

Mechanistic history

Synthesis pass · year-by-year structured walk · 23 steps
  1. 1994 High

    Established the identity of p67phox as the direct Rac effector in the oxidase, answering how the small GTPase signal is transduced to the enzyme.

    Evidence Direct binding assay with Rac effector-site mutagenesis and cell-free oxidase reconstitution

    PMID:8036496

    Open questions at the time
    • Did not define the structural basis of GTPase specificity
    • Did not distinguish Rac1 vs Rac2 preference
  2. 1996 Medium

    Resolved the Rac isoform preference and confirmed the p47phox–p67phox interaction, refining which partners assemble with p67phox.

    Evidence Yeast two-hybrid with multiple Rac effector-site mutants

    PMID:8550629

    Open questions at the time
    • Yeast two-hybrid only, no quantitative affinity
    • Functional consequence of preference not directly tested
  3. 1997 High

    Quantified the Rac–p67phox interaction and mapped the Rac effector region, separating binding from activation contributions.

    Evidence Fluorescent GTP-analog binding assay and Rac mutagenesis with superoxide kinetics

    PMID:9228059

    Open questions at the time
    • Identity of the partner contacted by the Rac insert region left unresolved
  4. 2000 High

    Provided the structural mechanism of GTPase specificity by showing the TPR insertion mediates the Rac·GTP interaction.

    Evidence X-ray crystallography of the TPR domain–Rac·GTP complex

    PMID:11090627

    Open questions at the time
    • Did not capture the activation domain or full-length context
    • Conformational coupling to activation not addressed
  5. 1999 High

    Defined the catalytic role of the activation domain by showing it controls hydride transfer from NADPH to FAD rather than NADPH binding, pinpointing the enzymatic step p67phox regulates.

    Evidence Fluorescent FAD-analog reconstitution, kinetic deuterium isotope effects, mutagenesis (building on the 1998 V204A activation-domain mapping)

    PMID:10438466 PMID:9642219

    Open questions at the time
    • Atomic contact between the activation domain and flavocytochrome b558 not defined
  6. 1995 High

    Dissected the division of labor between cytosolic factors, showing p67phox drives FAD reduction while p47phox enables electron flow beyond FAD.

    Evidence Cell-free reconstitution measuring FAD reduction with CGD cytosols deficient in p47phox or p67phox

    PMID:7896790

    Open questions at the time
    • Did not resolve the structural basis of the p47phox requirement for heme reduction
  7. 2002 High

    Established the architecture of the resting cytosolic complex, placing p67phox as the bridging molecule of a 1:1:1 heterotrimer.

    Evidence Isothermal titration calorimetry and gel filtration with interaction mapping

    PMID:11796733

    Open questions at the time
    • Conformational changes upon activation not captured
  8. 1998 High

    Identified the PC motif of p40phox as the module binding the inter-SH3 region of p67phox, defining the molecular basis of the p40phox–p67phox link.

    Evidence Yeast two-hybrid, in vitro binding, and PC-motif mutagenesis (extending earlier inter-SH3 mapping)

    PMID:7890694 PMID:9490029

    Open questions at the time
    • Functional consequence of disrupting the link in intact cells not measured here
  9. 1996 Medium

    Showed p67phox itself contains an NADPH-binding site contributing to catalysis, addressing where nucleotide engages the complex.

    Evidence NADPH dialdehyde affinity labeling and cell-free reconstitution with recombinant p67phox

    PMID:8770870

    Open questions at the time
    • Partially contradicted by later work assigning NADPH binding to gp91phox
    • Site not mapped at residue level
  10. 2004 High

    Defined Rac's dual function—membrane tethering plus induction of an activating conformational change in the activation domain—integrating binding and activation.

    Evidence p67phox(1-212)–Rac1 chimera mutagenesis with cell-free reconstitution and translocation assays

    PMID:14761978

    Open questions at the time
    • Conformational change inferred functionally, not visualized structurally
  11. 2001 High

    Provided high-resolution structure of the active N-terminal fragment and linked a CGD mutation to temperature-sensitive folding, connecting structure to disease.

    Evidence 1.8 Å crystal structure with temperature-dependent reconstitution and mutagenesis

    PMID:11262407

    Open questions at the time
    • Activation domain assignment within the C-terminal helix not definitively settled
  12. 1998 Medium

    Revealed autoinhibitory intramolecular regulation, showing Rac-binding sites are cryptic until C-terminal SH3/polyproline elements are released.

    Evidence In vitro binding with truncation mutants and PAK phosphorylation assay

    PMID:9624165

    Open questions at the time
    • Trigger that relieves autoinhibition in vivo not identified
    • Single lab
  13. 1999 High

    Identified Thr233 as the major phosphorylation site, anchoring the phospho-regulation of p67phox to a defined residue.

    Evidence 32P labeling, phosphopeptide mapping, HPLC-MS, and T233A mutagenesis (with earlier PKC in-cell/in-vitro mapping)

    PMID:9202043 PMID:9931304

    Open questions at the time
    • Functional consequence of Thr233 phosphorylation for oxidase activity not directly quantified
  14. 2003 High

    Assigned ERK2 and p38MAPK to distinct N- and C-terminal regions and revealed intramolecular masking of a phosphosite, refining the kinase regulation map.

    Evidence In vitro kinase assays with truncation mutants and inhibitor studies in neutrophils

    PMID:12693948

    Open questions at the time
    • How phosphorylation alters assembly or activity mechanistically not resolved
  15. 2004 High

    Identified PKCδ as the cell-type-specific kinase for p67phox in monocytes, linking a defined kinase to oxidase output.

    Evidence Co-IP, in vitro kinase assay, rottlerin/antisense inhibition, superoxide measurement

    PMID:15591124

    Open questions at the time
    • Specific PKCδ target residues on p67phox not mapped
  16. 1996 High

    Established the carrier hierarchy of translocation, showing p67phox mediates p40phox and Rac1 translocation while Rac2 moves independently.

    Evidence Subcellular fractionation of p47phox- and p67phox-deficient CGD neutrophils (with companion Rac1 translocation work)

    PMID:8120032 PMID:8670049 PMID:8948460

    Open questions at the time
    • Molecular interactions driving each translocation route not fully defined
  17. 2006 High

    Resolved that p67phox cannot translocate alone and uses p40phox versus p47phox as compartment-specific carriers to endosomes or plasma membrane.

    Evidence GFP-tagged live-cell imaging with PX-domain mutations and co-translocation assays

    PMID:17122360

    Open questions at the time
    • Physiological significance of distinct membrane compartments for oxidase function not delineated
  18. 1999 High

    Showed flavocytochrome b558 is required for stable phagosomal retention of p67phox, distinguishing transient recruitment from stable assembly.

    Evidence Synchronized phagocytosis with imaging and phagosomal fractionation in X-CGD cells (with FRAP-based exchange dynamics)

    PMID:10233905 PMID:14623873 PMID:7964505 PMID:8182143

    Open questions at the time
    • Molecular contacts stabilizing the membrane-bound complex not mapped at residue level
  19. 2005 High

    Identified S100A8/A9 as a positive regulator that delivers arachidonic acid and directly binds p67phox/Rac to promote oxidase activation, adding a lipid-mediated activation input.

    Evidence Co-IP, cell-free assay, S100A9 KO and sihRNA, arachidonate-binding mutant across multiple systems

    PMID:15642721

    Open questions at the time
    • Structural basis of the S100A8–p67phox interaction not determined
  20. 1999 Medium

    Defined the transcriptional control of NCF2, showing PU.1/IRF1/ICSBP recruit CBP at a specific cis element for IFN-γ-induced expression, with downstream layers of HoxA10 repression, SHP1 modulation, and p53 activation.

    Evidence Reporter assays, ChIP, transcription-factor manipulation, and tyrosine-phosphorylation mutagenesis across studies

    PMID:10570299 PMID:11483597 PMID:16210632 PMID:23187810

    Open questions at the time
    • Integration of these regulators during in vivo myeloid differentiation not fully reconstructed
    • Single-lab reporter-based mechanisms
  21. 2011 Medium

    Connected NCF2 variants to disease and signaling beyond CGD, with a PB1-domain residue implicated in Vav1 coupling and a RAC2-binding variant linked to very early onset IBD.

    Evidence Site-directed mutagenesis with ROS readouts, computational modeling, and patient variant binding/activity assays

    PMID:21900546 PMID:22203994 PMID:8879195

    Open questions at the time
    • Vav1 interaction predicted computationally, not directly demonstrated structurally
    • Causal mechanism linking reduced oxidase to IBD pathology not established
  22. 2015 Medium

    Extended p67phox into innate immune signaling, showing TLR/IRAK-ERK-driven p67phox–Nox2 assembly generates ROS required for IL-1β processing.

    Evidence siRNA knockdown, co-IP of ERK–p67phox and p67phox–Nox2, translocation and IL-1β readouts

    PMID:26320741

    Open questions at the time
    • Direct vs indirect nature of the ERK–p67phox interaction not established
    • Single lab
  23. 2019 High

    Demonstrated pathogen subversion of p67phox, showing M. tuberculosis PPE2 binds p67phox to block translocation and ROS production, aiding bacterial survival.

    Evidence Co-IP, Trp236Ala mutagenesis, translocation and ROS assays, bacterial survival in macrophages

    PMID:31375544

    Open questions at the time
    • Binding interface on p67phox not mapped
    • In vivo relevance during infection not tested

Open questions

Synthesis pass · forward-looking unresolved questions
  • How phosphorylation events, autoinhibition release, and Rac-induced conformational change are temporally coordinated to switch the resting heterotrimer into an active membrane-bound oxidase remains unresolved.
  • No integrated structure of the assembled active complex on flavocytochrome b558
  • Functional ordering of phospho-events relative to translocation undefined
  • Residue-level p67phox–NOX2 activation interface unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 3 GO:0098772 molecular function regulator activity 3 GO:0140096 catalytic activity, acting on a protein 3 GO:0008092 cytoskeletal protein binding 2
Localization
GO:0005829 cytosol 3 GO:0005886 plasma membrane 3 GO:0005768 endosome 1
Pathway
R-HSA-168256 Immune System 4 R-HSA-162582 Signal Transduction 3
Partners
CYBBMAPK1NCF1NCF4PRKCDRAC1RAC2S100A8
Complex memberships
p40phox-p47phox-p67phox cytosolic heterotrimerphagocyte NADPH oxidase

Evidence

Reading pass · 42 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 Rac1 interacts directly with p67phox in a GTP-dependent manner; effector-site mutations in Rac that abolished NADPH oxidase activity also abolished binding to p67phox, establishing p67phox as the Rac effector protein in the NADPH oxidase complex. Direct binding assay, functional cell-free oxidase reconstitution, Rac effector-site mutagenesis Science High 8036496
2000 Crystal structure of the N-terminal TPR domain of p67phox bound to Rac·GTP reveals a novel Rho-family/effector interaction mode; complex formation is largely mediated by an insertion between two TPR motifs, explaining GTPase specificity. X-ray crystallography Molecular Cell High 11090627
1998 An activation domain in p67phox spanning residues 199–210 is essential for NADPH oxidase activity; the V204A single-residue mutation completely abolishes superoxide generation without affecting Rac binding, and acts as a dominant-negative inhibitor, indicating direct activation of flavocytochrome b558 by this domain. C-terminal truncation mutagenesis, single-residue mutagenesis, cell-free reconstitution assay, competition/translocation experiments Journal of Biological Chemistry High 9642219
1999 The activation domain of p67phox (residues 199–210) regulates the reductive half-reaction of FAD in flavocytochrome b558 (hydride transfer from NADPH to FAD), not NADPH binding per se, as shown by fluorescent FAD analog reconstitution and kinetic deuterium isotope experiments. Fluorescent FAD analog (thioacetamido-FAD) reconstitution, steady-state flavin fluorescence, kinetic deuterium isotope effects, mutagenesis Journal of Biological Chemistry High 10438466
2009 SAXS analysis reveals that p67phox is an elongated multidomain protein with semi-flexible linkers organized as beads on a string, with evidence for intramolecular interactions; p67phox is the only NADPH oxidase cytosolic factor that interacts with all other components. Small-angle X-ray scattering (SAXS) Journal of Structural Biology Medium 19723583
2002 The cytosolic p40phox–p47phox–p67phox complex contains one copy of each protein (heterotrimeric), has an extended non-globular shape, and p67phox is the primary binding partner of p47phox in the resting state, acting as the bridging molecule connecting p40phox and p47phox. Isothermal titration calorimetry (ITC), gel filtration, detailed protein–protein interaction mapping Journal of Biological Chemistry High 11796733
1994 Both SH3 domains of p67phox are required for membrane binding and NADPH oxidase activity in whole cells; the N-terminal domain of p67phox (residues 1–246) lacking SH3 domains retains activity in cell-free assays but not in intact cells, indicating SH3 domains are dispensable for direct enzyme activation but essential for membrane targeting. SH3 domain deletion constructs, whole-cell NADPH oxidase activity assay in CGD B-cells, cell-free reconstitution Journal of Biological Chemistry High 8206939
1997 Rac1 and Rac2 bind p67phox directly with 1:1 stoichiometry (Kd ~120 nM and ~60 nM respectively); the Rac1 effector region (residues 26–45) mediates binding to p67phox, while the insert region (residues 124–135) affects activity without affecting p67phox binding affinity, suggesting interaction with another oxidase component. Fluorescent GTP analog (mant-GppNHp) binding assay, steady-state kinetics of superoxide generation, Rac mutagenesis Journal of Biological Chemistry High 9228059
1996 p67phox interacts preferentially with Rac2 over Rac1 (~6-fold) in GTP-dependent manner as shown by yeast two-hybrid; Rac effector-site mutants inactive in NADPH oxidase lose interaction with p67phox; a strong intracellular interaction between p47phox and p67phox was also confirmed. Yeast two-hybrid system Journal of Biological Chemistry Medium 8550629
1994 p40phox co-immunoprecipitates with p67phox in a 1:1 molar complex in resting neutrophils and is absent in CGD patients lacking p67phox, indicating a constitutive stoichiometric cytosolic complex. Co-immunoprecipitation, gel filtration, immunoblot Biochemical and Biophysical Research Communications Medium 8147882
1995 The C-terminal half of p40phox (but not its SH3 domain) interacts with p67phox; the interaction site on p67phox lies in the region between the two SH3 domains (not the SH3 domains themselves), representing a non-canonical protein–protein interaction module. Yeast two-hybrid system, in vitro binding assays with truncated constructs Journal of Biological Chemistry Medium 7890694
1998 A PC motif in the C-terminal region of p40phox is the critical module for binding to the region between the two SH3 domains of p67phox; site-directed mutagenesis of the PC motif abolished p40phox–p67phox interaction. Yeast two-hybrid, in vitro binding with recombinant proteins, site-directed mutagenesis European Journal of Biochemistry High 9490029
1997 p67phox is phosphorylated in human neutrophils upon stimulation with fMLP or PMA; PKC phosphorylates p67phox in vitro on the same peptide as in intact cells; fMLP-induced phosphorylation is only minimally affected by PKC inhibition, indicating PKC-dependent and PKC-independent phosphorylation pathways. Immunoprecipitation, 32P metabolic labeling, two-dimensional tryptic peptide mapping, in vitro kinase assay with recombinant GST-p67phox Journal of Biological Chemistry High 9202043
2003 ERK2 phosphorylates the N-terminal fragment (residues 1–243) of p67phox while p38MAPK phosphorylates the C-terminal fragment (residues 244–526) at selective sites; an N-terminal fragment containing the TPR-rich region masks a C-terminal phosphorylation site, revealing an intramolecular regulatory domain. Both kinases phosphorylate p67phox in intact fMLP- and PMA-stimulated neutrophils. In vitro kinase assays with truncation mutants, two-dimensional phosphopeptide mapping, MEK1/2 and p38MAPK inhibitors in intact neutrophils Biochemistry High 12693948
1999 The major phosphorylation site of p67phox is Thr233, located in the proline-rich domain; mutagenesis of Thr233 to alanine abolished in vitro phosphorylation by both cytosol and MAP kinase. Metabolic 32P labeling, phosphopeptide mapping, HPLC-MS, site-directed mutagenesis (T233A) Biochemical Journal High 9931304
1995 p67phox alone facilitates electron flow from NADPH to the flavin (FAD) of NADPH oxidase independent of p47phox; p47phox is required for electron transfer to proceed beyond FAD to heme in cytochrome b-245 and thence to oxygen, establishing distinct roles for each cytosolic activating factor. Cell-free reconstitution assay measuring FAD reduction and dye reductase activity, using CGD cytosols deficient in p47phox or p67phox Journal of Biological Chemistry High 7896790
1996 p67phox contains an NADPH-binding site that participates in catalysis: treatment of cytosol with NADPH dialdehyde inactivates oxidase function, and this inactivation is reversed by recombinant p67phox; purified recombinant p67phox binds [32P]NADPH dialdehyde specifically and its cell-free activity is abolished by NADPH dialdehyde treatment. Affinity labeling with NADPH dialdehyde, cell-free oxidase reconstitution, [32P] NADPH dialdehyde binding to purified recombinant p67phox Journal of Clinical Investigation Medium 8770870
1993 Translocation of p67phox to the plasma membrane and its phosphorylation are correlated with NADPH oxidase activation; continuous phosphorylation and translocation of p67phox is required to maintain activated oxidase activity, established using p67phox-deficient CGD neutrophils as controls. Immunoprecipitation from subcellular fractions, 32P metabolic labeling, comparison with p67phox-deficient CGD cells Biochemical Journal High 8257426
1994 Rac2 translocation to the submembranous actin cytoskeleton upon neutrophil activation is dependent on p47phox but not on p67phox; p67phox, Rac2, and NADPH oxidase activity are restricted to the membrane cytoskeleton in activated cells. Subcellular fractionation of normal and p47phox-deficient CGD neutrophils, immunoblot Journal of Biological Chemistry Medium 8120032
1996 In p67phox-deficient CGD neutrophils, p47phox and Rac2 still translocate upon stimulation, but p40phox and Rac1 do not; conversely, in p47phox-deficient neutrophils, p67phox, p40phox, and Rac1 fail to translocate. This establishes that p67phox mediates translocation of p40phox and Rac1 while Rac2 translocates independently. Subcellular fractionation of CGD patient neutrophils (p47phox-deficient and p67phox-deficient), immunoblot Biochemical Journal High 8670049
1995 In p67phox-deficient CGD neutrophils, Rac1 fails to translocate to membranes upon stimulation, establishing that p67phox is required for Rac1 (but not Rac2) translocation; an interaction between p67phox and Rac1 is essential for translocation of cytosolic proteins and NADPH oxidase activation. Subcellular fractionation of p67phox-deficient CGD patient neutrophils, immunoblot Biochemical Journal Medium 8948460
1996 A CGD patient with an in-frame deletion of Lys58 in p67phox produces a nonfunctional protein that cannot interact with Rac1, and p47phox and p67phox fail to translocate to the membrane upon activation; this is the first disease-causing mutation attributable to disrupted Rac/effector interaction. Patient genetic analysis (cDNA/genomic sequencing), co-immunoprecipitation, functional neutrophil activation assays Journal of Experimental Medicine High 8879195
2004 A Rac-p67phox chimera analysis demonstrates Rac has a dual role in NADPH oxidase assembly: (1) tethering p67phox to the membrane via the Rac C-terminal polybasic region, and (2) inducing an activating conformational change in the p67phox activation domain via the intrachimeric p67phox–Rac1 interaction. Construction and mutagenesis of p67phox-(1-212)-Rac1 chimeras, cell-free NADPH oxidase reconstitution, competitive inhibition and translocation assays Journal of Biological Chemistry High 14761978
1998 Rac-binding sites of p67phox (residues 170–199) are cryptic in the full-length protein; deletion of the C-terminal SH3 domain or polyproline motif unmasks Rac1/2 binding ~8-fold and also exposes a PAK phosphorylation site adjacent to the Rac-binding domain, suggesting autoinhibitory intramolecular regulation. In vitro binding assays with p67phox truncation/deletion mutants, PAK kinase phosphorylation assay Journal of Biological Chemistry Medium 9624165
2001 Crystal structure of the active N-terminal fragment of p67phox (residues 1–213) at 1.8 Å resolution reveals four TPR motifs with the C-terminus folding into a hydrophobic groove; a short C-terminal helix (residues 187–193) may be part of the activation domain. A CGD mutation A128V causes temperature-sensitive loss of function despite full in vitro activity at 25°C. X-ray crystallography, functional cell-free reconstitution at different temperatures, mutagenesis (G78E, A128V) Journal of Biological Chemistry High 11262407
2006 p67phox does not translocate to membranes when expressed alone; it depends on p40phox for translocation to early endosomes or on p47phox for translocation to the plasma membrane, establishing p40phox and p47phox as distinct carrier proteins for p67phox targeting to different membrane compartments. GFP-tagged phox protein live-cell imaging, PX domain mutations disrupting phospholipid binding, co-translocation assays in cell lines Molecular Biology of the Cell High 17122360
1994 p47phox and p67phox membrane translocation requires intact cytochrome b558 (gp91phox/p22phox) for stable association; a mutation at p22phox Pro156Gln virtually abolishes translocation of both p47phox and p67phox without affecting intrinsic cytochrome b558 electron flow capacity. Cell-free translocation assay, immunoblot of subcellular fractions from CGD patient neutrophils Journal of Experimental Medicine High 7964505
1994 A mutation at Asp500Gly in gp91phox is associated with normal cytochrome b558 levels but strongly disrupts translocation of both p47phox and p67phox to the membrane; a synthetic peptide mimicking gp91phox domain 491–504 inhibits both NADPH oxidase activity and p47phox/p67phox translocation in cell-free assays. Cell-free translocation assay, synthetic peptide inhibition, immunoblot Journal of Clinical Investigation Medium 8182143
2003 FRAP analysis of GFP-p67phox on phagosomes in living PLB-985 cells shows a continuous, rapid exchange of membrane-bound p67phox with cytosolic p67phox during phagocytosis; this exchange does not depend on actin cytoskeleton rearrangement. In X-CGD cells lacking flavocytochrome b558, p67phox is transiently recruited but fails to be stably retained. GFP-tagged protein live-cell imaging, fluorescence recovery after photobleaching (FRAP), cytochalasin B treatment Journal of Biological Chemistry High 14623873
1999 In CGD neutrophils lacking flavocytochrome b558 (X-linked CGD), p47phox and p67phox are transiently recruited to phagosomes but are shed from phagosomal membranes along with F-actin once phagocytosis is complete, unlike in normal cells where they are stably retained; this establishes flavocytochrome b558 as required for stable phagosomal membrane binding of p47phox and p67phox. Synchronized phagocytosis assay, immunofluorescence microscopy, immunoblotting of phagosomal fractions Blood High 10233905
2005 S100A8 (but not S100A9) directly binds p67phox and Rac, and the S100A8/A9 complex promotes NADPH oxidase activation by transferring arachidonic acid to the complex; an arachidonic acid-binding mutant of S100A8/A9 fails to enhance oxidase activity. Co-immunoprecipitation, cell-free oxidase assay, S100A9 knockout mouse neutrophils, S100A9 siRNA in NB4 cells, arachidonate-binding mutant FASEB Journal High 15642721
2008 PKC phosphorylation of gp91phox/NOX2 enhances its binding to p67phox (as well as to Rac2 and p47phox) and increases diaphorase activity, representing a novel regulatory mechanism for NADPH oxidase complex assembly. In vitro PKC phosphorylation of recombinant gp91phox cytosolic domain, two-dimensional tryptic peptide mapping, binding assay, diaphorase activity assay FASEB Journal Medium 19028840
2004 PKCδ forms a complex with p67phox in activated monocytes, phosphorylates p67phox in vitro, and PKCδ inhibition (rottlerin or antisense) blocks p67phox phosphorylation and reduces NADPH oxidase-dependent superoxide production, establishing PKCδ as a specific kinase for p67phox in primary human monocytes. Co-immunoprecipitation (PKCδ-p67phox complex), in vitro kinase assay, rottlerin/antisense PKCδ inhibition, superoxide measurement Journal of Leukocyte Biology High 15591124
1999 Recruitment of CREB-binding protein (CBP) by the combined action of PU.1, IRF1, and ICSBP to the NCF2 promoter is the molecular mechanism of IFN-γ-induced p67phox transcription; a specific NCF2 cis element is necessary and sufficient for this activation. Reporter gene (luciferase) assays, transcription factor overexpression, mutational analysis of NCF2 cis elements Journal of Immunology Medium 10570299
2001 SHP1 protein-tyrosine phosphatase inhibits NCF2/p67phox expression in undifferentiated myeloid cells by decreasing the interaction of PU.1, IRF1, ICSBP, and CBP with the NCF2 cis element; tyrosine phosphorylation of IRF1 and ICSBP is required for stage-specific NCF2 transcription during differentiation. Reporter gene assays, chromatin immunoprecipitation, SHP1 overexpression, site-directed mutagenesis of IRF1/ICSBP tyrosine residues Journal of Biological Chemistry Medium 11483597
2005 HoxA10 represses NCF2/p67phox transcription in undifferentiated myeloid cells via a homologous NCF2 promoter cis element; repression requires histone deacetylase activity and is relieved by IFN-γ-induced tyrosine phosphorylation of HoxA10's homeodomain. Reporter gene assay, chromatin immunoprecipitation, HDAC inhibitor treatment, IFN-γ stimulation, mutagenesis of HoxA10 tyrosines Journal of Immunology Medium 16210632
2012 p53 directly binds the NCF2 promoter in vivo and activates NCF2/p67phox transcription; NCF2 knockdown reduces ROS production and promotes cell death, suggesting Nox2-generated ROS have a protective function against apoptosis downstream of p53. Chromatin immunoprecipitation (ChIP), luciferase reporter assay, siRNA knockdown, RT-PCR across multiple p53-inducible cell lines Cell Cycle Medium 23187810
2011 The NCF2 H389Q missense mutation (in the PB1 domain) reduces NADPH oxidase-dependent ROS production ~2-fold in Vav-dependent FcγR-elicited activation; computational modeling predicts H389 interacts with Vav1 ZF domain residues and H389Q weakens binding by ~1.5 kcal/mol. Site-directed mutagenesis at position 389, ROS measurement in FcγR-stimulated cells, computational (molecular dynamics) modeling PNAS Medium 22203994
2011 A rare NCF2 missense variant reduces binding of p67phox to RAC2 and is associated with very early onset IBD; the reduced RAC2 binding results in partial inhibition of NADPH oxidase function. Direct sequencing, functional binding assay (p67phox–RAC2 interaction), NADPH oxidase activity assay in patient cells Gut Medium 21900546
2019 Mycobacterium tuberculosis PPE2 protein directly interacts with p67phox via its SH3-like domain (Trp236 is critical); PPE2 prevents translocation of p67phox and p47phox to the membrane, inhibiting NADPH oxidase-mediated ROS production and enhancing mycobacterial survival in macrophages. Co-immunoprecipitation, Trp236Ala mutagenesis of PPE2, p67phox/p47phox translocation assay, ROS measurement, bacterial survival assay in macrophages Journal of Immunology High 31375544
2015 TLR4/TLR2 signaling causes IRAK-dependent ERK-p67phox interaction and p67phox translocation; the resulting p67phox–Nox-2 interaction drives ROS generation required for IL-1β transcription and caspase-1-dependent processing in monocytes. siRNA knockdown of Nox-2 and IRAK, co-immunoprecipitation of ERK-p67phox and p67phox-Nox-2, p67phox translocation assay, ROS measurement, IL-1β quantification Cellular & Molecular Immunology Medium 26320741
2003 PKC-β activation is required for translocation of p47phox and p67phox from cytoplasm to membrane in diabetic glomeruli; adenoviral PKC-β2 overexpression in mesangial cells enhances ROS generation by promoting this translocation, and the selective PKC-β inhibitor ruboxistaurin prevents it. Subcellular fractionation immunoblot, adenoviral PKC-β2 overexpression, PKC-β inhibitor treatment, superoxide measurement in diabetic rat glomeruli and mesangial cells Diabetes Medium 14514646

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1994 Interaction of Rac with p67phox and regulation of phagocytic NADPH oxidase activity. Science (New York, N.Y.) 340 8036496
2003 Novel human homologues of p47phox and p67phox participate in activation of superoxide-producing NADPH oxidases. The Journal of biological chemistry 316 12716910
2000 Structure of the TPR domain of p67phox in complex with Rac.GTP. Molecular cell 261 11090627
2003 Proteins homologous to p47phox and p67phox support superoxide production by NAD(P)H oxidase 1 in colon epithelial cells. The Journal of biological chemistry 241 12657628
2009 NADPH oxidase activator p67(phox) behaves in solution as a multidomain protein with semi-flexible linkers. Journal of structural biology 240 19723583
1994 Rac translocates independently of the neutrophil NADPH oxidase components p47phox and p67phox. Evidence for its interaction with flavocytochrome b558. The Journal of biological chemistry 209 7982999
1998 Angiotensin II induces p67phox mRNA expression and NADPH oxidase superoxide generation in rabbit aortic adventitial fibroblasts. Hypertension (Dallas, Tex. : 1979) 192 9719063
1998 Regulation of the neutrophil respiratory burst oxidase. Identification of an activation domain in p67(phox). The Journal of biological chemistry 188 9642219
2003 Translocation of glomerular p47phox and p67phox by protein kinase C-beta activation is required for oxidative stress in diabetic nephropathy. Diabetes 182 14514646
1994 Cytosolic guanine nucleotide-binding protein Rac2 operates in vivo as a component of the neutrophil respiratory burst oxidase. Transfer of Rac2 and the cytosolic oxidase components p47phox and p67phox to the submembranous actin cytoskeleton during oxidase activation. The Journal of biological chemistry 167 8120032
2000 Upregulation of p67(phox) and gp91(phox) in aortas from angiotensin II-infused mice. American journal of physiology. Heart and circulatory physiology 151 11045958
2011 Lupus-associated causal mutation in neutrophil cytosolic factor 2 (NCF2) brings unique insights to the structure and function of NADPH oxidase. Proceedings of the National Academy of Sciences of the United States of America 150 22203994
2005 The arachidonic acid-binding protein S100A8/A9 promotes NADPH oxidase activation by interaction with p67phox and Rac-2. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 149 15642721
2011 NADPH oxidase complex and IBD candidate gene studies: identification of a rare variant in NCF2 that results in reduced binding to RAC2. Gut 148 21900546
1999 The p67(phox) activation domain regulates electron flow from NADPH to flavin in flavocytochrome b(558). The Journal of biological chemistry 148 10438466
2006 A p67Phox-like regulator is recruited to control hyphal branching in a fungal-grass mutualistic symbiosis. The Plant cell 140 17041146
2008 Regulation of the phagocyte NADPH oxidase activity: phosphorylation of gp91phox/NOX2 by protein kinase C enhances its diaphorase activity and binding to Rac2, p67phox, and p47phox. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 139 19028840
1995 The expression of NADPH oxidase components in human glomerular mesangial cells: detection of protein and mRNA for p47phox, p67phox, and p22phox. Journal of the American Society of Nephrology : JASN 133 7703387
2002 Architecture of the p40-p47-p67phox complex in the resting state of the NADPH oxidase. A central role for p67phox. The Journal of biological chemistry 131 11796733
2004 Nox3 regulation by NOXO1, p47phox, and p67phox. The Journal of biological chemistry 129 15181005
2012 Increased expression of NAD(P)H oxidase subunit p67(phox) in the renal medulla contributes to excess oxidative stress and salt-sensitive hypertension. Cell metabolism 123 22326221
1997 Phosphorylation of the respiratory burst oxidase subunit p67(phox) during human neutrophil activation. Regulation by protein kinase C-dependent and independent pathways. The Journal of biological chemistry 122 9202043
2018 LINC01410-miR-532-NCF2-NF-kB feedback loop promotes gastric cancer angiogenesis and metastasis. Oncogene 121 29483646
1991 Characterization of neutrophil NADPH oxidase factors p47-phox and p67-phox from recombinant baculoviruses. The Journal of biological chemistry 119 1918085
2004 Dual role of Rac in the assembly of NADPH oxidase, tethering to the membrane and activation of p67phox: a study based on mutagenesis of p67phox-Rac1 chimeras. The Journal of biological chemistry 112 14761978
1997 Rac binding to p67(phox). Structural basis for interactions of the Rac1 effector region and insert region with components of the respiratory burst oxidase. The Journal of biological chemistry 111 9228059
1993 Relationship between phosphorylation and translocation to the plasma membrane of p47phox and p67phox and activation of the NADPH oxidase in normal and Ca(2+)-depleted human neutrophils. The Biochemical journal 111 8439286
1996 Mechanisms of NADPH oxidase activation: translocation of p40phox, Rac1 and Rac2 from the cytosol to the membranes in human neutrophils lacking p47phox or p67phox. The Biochemical journal 108 8670049
1999 Recruitment of CREB-binding protein by PU.1, IFN-regulatory factor-1, and the IFN consensus sequence-binding protein is necessary for IFN-gamma-induced p67phox and gp91phox expression. Journal of immunology (Baltimore, Md. : 1950) 101 10570299
1995 Mapping the domains of interaction of p40phox with both p47phox and p67phox of the neutrophil oxidase complex using the two-hybrid system. The Journal of biological chemistry 101 7890694
2003 Phosphorylation of the NADPH oxidase component p67(PHOX) by ERK2 and P38MAPK: selectivity of phosphorylated sites and existence of an intramolecular regulatory domain in the tetratricopeptide-rich region. Biochemistry 95 12693948
1994 A novel cytosolic component, p40phox, of respiratory burst oxidase associates with p67phox and is absent in patients with chronic granulomatous disease who lack p67phox. Biochemical and biophysical research communications 95 8147882
1996 The Rac target NADPH oxidase p67phox interacts preferentially with Rac2 rather than Rac1. The Journal of biological chemistry 92 8550629
1994 156Pro-->Gln substitution in the light chain of cytochrome b558 of the human NADPH oxidase (p22-phox) leads to defective translocation of the cytosolic proteins p47-phox and p67-phox. The Journal of experimental medicine 91 7964505
1994 Role of p67-phox SH3 domains in assembly of the NADPH oxidase system. The Journal of biological chemistry 91 8206939
1995 The cytosolic activating factors p47phox and p67phox have distinct roles in the regulation of electron flow in NADPH oxidase. The Journal of biological chemistry 90 7896790
1994 A point mutation in gp91-phox of cytochrome b558 of the human NADPH oxidase leading to defective translocation of the cytosolic proteins p47-phox and p67-phox. The Journal of clinical investigation 89 8182143
1993 Activation of NADPH oxidase of human neutrophils involves the phosphorylation and the translocation of cytosolic p67phox. The Biochemical journal 89 8257426
2002 Cytosolic phospholipase A2 (cPLA2) regulation of human monocyte NADPH oxidase activity. cPLA2 affects translocation but not phosphorylation of p67(phox) and p47(phox). The Journal of biological chemistry 86 12101222
2006 Homocysteine stimulates phosphorylation of NADPH oxidase p47phox and p67phox subunits in monocytes via protein kinase Cbeta activation. The Biochemical journal 81 16626305
1999 Transient association of the nicotinamide adenine dinucleotide phosphate oxidase subunits p47phox and p67phox with phagosomes in neutrophils from patients with X-linked chronic granulomatous disease. Blood 79 10233905
1995 Dissociation of Rac translocation from p47phox/p67phox movements in human neutrophils by tyrosine kinase inhibitors. Journal of leukocyte biology 77 7616102
2006 A regulated adaptor function of p40phox: distinct p67phox membrane targeting by p40phox and by p47phox. Molecular biology of the cell 75 17122360
1995 A 68-kDa kinase and NADPH oxidase component p67phox are targets for Cdc42Hs and Rac1 in neutrophils. The Journal of biological chemistry 75 7738010
1996 Interactions between cytosolic components of the NADPH oxidase: p40phox interacts with both p67phox and p47phox. The Biochemical journal 73 8760383
1998 Anionic amphiphile-independent activation of the phagocyte NADPH oxidase in a cell-free system by p47phox and p67phox, both in C terminally truncated forms. Implication for regulatory Src homology 3 domain-mediated interactions. The Journal of biological chemistry 71 9461621
1996 Involvement of p40phox in activation of phagocyte NADPH oxidase through association of its carboxyl-terminal, but not its amino-terminal, with p67phox. The Journal of experimental medicine 71 9064349
1998 The PC motif: a novel and evolutionarily conserved sequence involved in interaction between p40phox and p67phox, SH3 domain-containing cytosolic factors of the phagocyte NADPH oxidase. European journal of biochemistry 70 9490029
1998 Functional modules and expression of mouse p40(phox) and p67(phox), SH3-domain-containing proteins involved in the phagocyte NADPH oxidase complex. European journal of biochemistry 67 9490028
1995 Interactions between the cytosolic components p47phox and p67phox of the human neutrophil NADPH oxidase that are not required for activation in the cell-free system. The Journal of biological chemistry 67 7744754
1990 Genes for two autosomal recessive forms of chronic granulomatous disease assigned to 1q25 (NCF2) and 7q11.23 (NCF1). American journal of human genetics 67 2393022
1998 Cryptic Rac-binding and p21(Cdc42Hs/Rac)-activated kinase phosphorylation sites of NADPH oxidase component p67(phox). The Journal of biological chemistry 66 9624165
1994 The functional expression of p47-phox and p67-phox may contribute to the generation of superoxide by an NADPH oxidase-like system in human fibroblasts. FEBS letters 65 7982496
2013 Allelic heterogeneity in NCF2 associated with systemic lupus erythematosus (SLE) susceptibility across four ethnic populations. Human molecular genetics 64 24163247
2005 Effects of p47phox C terminus phosphorylations on binding interactions with p40phox and p67phox. Structural and functional comparison of p40phox and p67phox SH3 domains. The Journal of biological chemistry 64 15657040
1996 Disturbed interaction of p21-rac with mutated p67-phox causes chronic granulomatous disease. The Journal of experimental medicine 64 8879195
2001 The active N-terminal region of p67phox. Structure at 1.8 A resolution and biochemical characterizations of the A128V mutant implicated in chronic granulomatous disease. The Journal of biological chemistry 62 11262407
2002 A prenylated p67phox-Rac1 chimera elicits NADPH-dependent superoxide production by phagocyte membranes in the absence of an activator and of p47phox: conversion of a pagan NADPH oxidase to monotheism. The Journal of biological chemistry 60 11896062
2015 The IRAK-ERK-p67phox-Nox-2 axis mediates TLR4, 2-induced ROS production for IL-1β transcription and processing in monocytes. Cellular & molecular immunology 56 26320741
2008 Heme oxygenase-1 protects against neutrophil-mediated intestinal damage by down-regulation of neutrophil p47phox and p67phox activity and O2- production in a two-hit model of alcohol intoxication and burn injury. Journal of immunology (Baltimore, Md. : 1950) 56 18453614
2003 Continuous translocation of Rac2 and the NADPH oxidase component p67(phox) during phagocytosis. The Journal of biological chemistry 56 14623873
1994 Evidence for a readily dissociable complex of p47phox and p67phox in cytosol of unstimulated human neutrophils. The Journal of biological chemistry 56 8071369
2012 Identification of NCF2/p67phox as a novel p53 target gene. Cell cycle (Georgetown, Tex.) 55 23187810
2001 Activation of the superoxide-generating NADPH oxidase by chimeric proteins consisting of segments of the cytosolic component p67(phox) and the small GTPase Rac1. Biochemistry 55 11724569
2004 Protein kinase Cdelta regulates p67phox phosphorylation in human monocytes. Journal of leukocyte biology 54 15591124
2001 SHP1 protein-tyrosine phosphatase inhibits gp91PHOX and p67PHOX expression by inhibiting interaction of PU.1, IRF1, interferon consensus sequence-binding protein, and CREB-binding protein with homologous Cis elements in the CYBB and NCF2 genes. The Journal of biological chemistry 53 11483597
1999 Molecular characterization of autosomal recessive chronic granulomatous disease caused by a defect of the nicotinamide adenine dinucleotide phosphate (reduced form) oxidase component p67-phox. Blood 48 10498624
1996 Topological organization of the cytosolic activating complex of the superoxide-generating NADPH-oxidase. Pinpointing the sites of interaction between p47phoz, p67phox and p40phox using the two-hybrid system. Biochimica et biophysica acta 47 8679714
1999 P40phox associates with the neutrophil Triton X-100-insoluble cytoskeletal fraction and PMA-activated membrane skeleton: a comparative study with P67phox and P47phox. Journal of leukocyte biology 46 10614785
1994 p67-phox enhances the binding of p47-phox to the human neutrophil respiratory burst oxidase complex. The Journal of biological chemistry 46 8071333
2019 Mycobacterium tuberculosis PPE2 Protein Interacts with p67phox and Inhibits Reactive Oxygen Species Production. Journal of immunology (Baltimore, Md. : 1950) 44 31375544
1995 Mechanisms of NADPH oxidase activation in human neutrophils: p67phox is required for the translocation of rac 1 but not of rac 2 from cytosol to the membranes. The Biochemical journal 43 8948460
2006 Translocation of proteins homologous to human neutrophil p47phox and p67phox to the cell membrane in activated hemocytes of Galleria mellonella. Developmental and comparative immunology 41 16920193
2005 Impaired NADPH oxidase activity in Rac2-deficient murine neutrophils does not result from defective translocation of p47phox and p67phox and can be rescued by exogenous arachidonic acid. Journal of leukocyte biology 40 16275890
1996 The cytosolic subunit p67phox contains an NADPH-binding site that participates in catalysis by the leukocyte NADPH oxidase. The Journal of clinical investigation 40 8770870
1996 Neutrophils from patients after burn injury express a deficiency of the oxidase components p47-phox and p67-phox. Blood 40 8943869
2023 M6A-mediated-upregulation of lncRNA BLACAT3 promotes bladder cancer angiogenesis and hematogenous metastasis through YBX3 nuclear shuttling and enhancing NCF2 transcription. Oncogene 39 37612524
2001 Fused p47phox and p67phox truncations efficiently reconstitute NADPH oxidase with higher activity and stability than the individual components. The Journal of biological chemistry 39 11333262
2002 A 29-kDa protein associated with p67phox expresses both peroxiredoxin and phospholipase A2 activity and enhances superoxide anion production by a cell-free system of NADPH oxidase activity. The Journal of biological chemistry 38 12121978
1995 Inhibitory effect of porcine surfactant on the respiratory burst oxidase in human neutrophils. Attenuation of p47phox and p67phox membrane translocation as the mechanism. The Journal of clinical investigation 38 8675631
1992 Reconstitution and characterization of the human neutrophil respiratory burst oxidase using recombinant p47-phox, p67-phox and plasma membrane. Biochemical and biophysical research communications 38 1321612
1999 Autosomal recessive chronic granulomatous disease caused by novel mutations in NCF-2, the gene encoding the p67-phox component of phagocyte NADPH oxidase. Human genetics 37 10598813
2006 Activation of SHP2 protein-tyrosine phosphatase increases HoxA10-induced repression of the genes encoding gp91(PHOX) and p67(PHOX). The Journal of biological chemistry 36 17138561
1999 Rac1 disrupts p67phox/p40phox binding: a novel role for Rac in NADPH oxidase activation. Biochemical and biophysical research communications 36 10486263
2001 A fusion protein between rac and p67phox (1-210) reconstitutes NADPH oxidase with higher activity and stability than the individual components. Biochemistry 35 11705402
1997 Roles for proline-rich regions of p47phox and p67phox in the phagocyte NADPH oxidase activation in vitro. Biochemical and biophysical research communications 35 9425254
1997 Genetic correction of p67phox deficient chronic granulomatous disease using peripheral blood progenitor cells as a target for retrovirus mediated gene transfer. Blood 34 9057660
2010 A prenylated p47phox-p67phox-Rac1 chimera is a Quintessential NADPH oxidase activator: membrane association and functional capacity. The Journal of biological chemistry 33 20529851
2001 Modulation of microglial superoxide production by alpha-tocopherol in vitro: attenuation of p67(phox) translocation by a protein phosphatase-dependent pathway. Journal of neurochemistry 33 11752058
1993 Characterization of the p67phox gene: genomic organization and restriction fragment length polymorphism analysis for prenatal diagnosis in chronic granulomatous disease. Blood 33 7903171
2008 Platelet-activating factor-mediated endosome formation causes membrane translocation of p67phox and p40phox that requires recruitment and activation of p38 MAPK, Rab5a, and phosphatidylinositol 3-kinase in human neutrophils. Journal of immunology (Baltimore, Md. : 1950) 32 18523285
1999 The major phosphorylation site of the NADPH oxidase component p67phox is Thr233. The Biochemical journal 32 9931304
2016 Role of Nox4 and p67phox subunit of Nox2 in ROS production in response to increased tubular flow in the mTAL of Dahl salt-sensitive rats. American journal of physiology. Renal physiology 31 27279484
2014 Null mutation of the nicotinamide adenine dinucleotide phosphate-oxidase subunit p67phox protects the Dahl-S rat from salt-induced reductions in medullary blood flow and glomerular filtration rate. Hypertension (Dallas, Tex. : 1979) 29 25489057
2011 The NADPH oxidase cytosolic component p67phox is constitutively phosphorylated in human neutrophils: Regulation by a protein tyrosine kinase, MEK1/2 and phosphatases 1/2A. Biochemical pharmacology 29 21784060
2007 Tripartite chimeras comprising functional domains derived from the cytosolic NADPH oxidase components p47phox, p67phox, and Rac1 elicit activator-independent superoxide production by phagocyte membranes: an essential role for anionic membrane phospholipids. The Journal of biological chemistry 29 17548354
2005 HoxA10 represses transcription of the gene encoding p67phox in phagocytic cells. Journal of immunology (Baltimore, Md. : 1950) 29 16210632
2005 Expression of NOX-I, gp91phox, p47phox and P67phox in the aorta segments above and below coarctation. Biochimica et biophysica acta 28 15814300
1995 A mutation located at the 5' splice junction sequence of intron 3 in the p67phox gene causes the lack of p67phox mRNA in a patient with chronic granulomatous disease. Blood 27 7803798
2013 Evolutionary dynamics of the human NADPH oxidase genes CYBB, CYBA, NCF2, and NCF4: functional implications. Molecular biology and evolution 26 23821607

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