Molecular evolution of Phox-related regulatory subunits for NADPH oxidase enzymes
© Kawahara and Lambeth; licensee BioMed Central Ltd. 2007
Received: 16 June 2007
Accepted: 27 September 2007
Published: 27 September 2007
The reactive oxygen-generating N ADPH ox idases (Noxes) function in a variety of biological roles, and can be broadly classified into those that are regulated by subunit interactions and those that are regulated by calcium. The prototypical subunit-regulated Nox, Nox2, is the membrane-associated catalytic subunit of the phagocyte NADPH-oxidase. Nox2 forms a heterodimer with the integral membrane protein, p22phox, and this heterodimer binds to the regulatory subunits p47phox, p67phox, p40phox and the small GTPase Rac, triggering superoxide generation. Nox-organizer protein 1 (NOXO1) and Nox-activator 1 (NOXA1), respective homologs of p47phox and p67phox, together with p22phox and Rac, activate Nox1, a non-phagocytic homolog of Nox2. NOXO1 and p22phox also regulate Nox3, whereas Nox4 requires only p22phox. In this study, we have assembled and analyzed amino acid sequences of Nox regulatory subunit orthologs from vertebrates, a urochordate, an echinoderm, a mollusc, a cnidarian, a choanoflagellate, fungi and a slime mold amoeba to investigate the evolutionary history of these subunits.
Ancestral p47phox, p67phox, and p22phox genes are broadly seen in the metazoa, except for the ecdysozoans. The choanoflagellate Monosiga brevicollis, the unicellular organism that is the closest relatives of multicellular animals, encodes early prototypes of p22phox, p47phox as well as the earliest known Nox2-like ancestor of the Nox1-3 subfamily. p67phox- and p47phox-like genes are seen in the sea urchin Strongylocentrotus purpuratus and the limpet Lottia gigantea that also possess Nox2-like co-orthologs of vertebrate Nox1-3. Duplication of primordial p47phox and p67phox genes occurred in vertebrates, with the duplicated branches evolving into NOXO1 and NOXA1. Analysis of characteristic domains of regulatory subunits suggests a novel view of the evolution of Nox: in fish, p40phox participated in regulating both Nox1 and Nox2, but after the appearance of mammals, Nox1 (but not Nox2) became independent of p40phox. In the fish Oryzias latipes, a NOXO1 ortholog retains an autoinhibitory region that is characteristic of mammalian p47phox, and this was subsequently lost from NOXO1 in later vertebrates. Detailed amino acid sequence comparisons identified both putative key residues conserved in characteristic domains and previously unidentified conserved regions. Also, candidate organizer/activator proteins in fungi and amoeba are identified and hypothetical activation models are suggested.
This is the first report to provide the comprehensive view of the molecular evolution of regulatory subunits for Nox enzymes. This approach provides clues for understanding the evolution of biochemical and physiological functions for regulatory-subunit-dependent Nox enzymes.
Nox enzymes (reactive oxygen-generating NADPH-oxidases) diverged early in evolution into calcium-regulated Noxes (e.g., Nox5 and the Duox enzymes) and Noxes that are activated by binding to regulatory subunits [1, 2]. The most extensively studied of the latter is the ph agocyte NAPDH-ox idase (Phox), whose role in host defense has been documented at length [3–8]. Professional phagocytes such as neutrophils and macrophages produce large amounts of superoxide, with secondary production of other microbicidal reactive oxygen species (ROS). Superoxide is generated by the Phox, which consists of the catalytic subunit gp91phox (a.k.a. Nox2), along with the regulatory subunits p22phox, p67phox, p47phox, p40phox, and the small GTPase, Rac [3, 6, 9–11]. The importance of the oxidase is demonstrated by the inherited condition, chronic granulomatous disease (CGD), in which absent or mutated Phox proteins result in an inability of phagocytes to kill microbes [4, 12, 13].
Nox2 and p22phox are integral membrane proteins that form a heterodimer referred to as flavocytochrome b558. In resting cells, flavocytochrome b 558 is inactive and p47phox, p67phox, and p40phox are all present in the cytoplasm. Cell activation is accompanied by phosphorylation of p47phox and probably other Phox-regulatory subunits [3, 10, 14–16], and by guanine nucleotide exchange factors that convert Rac-GDP to Rac-GTP [3, 17]. These events trigger assembly of these proteins at the membrane, resulting in activation of Nox2. An intricate set of protein-protein interactions facilitates the binding of the regulatory subunits to the membrane components and to each other, and these interactions are mediated by well-characterized modular interaction domains. For example, p47phox and p67phox bind via a C-terminal proline-rich region (PRR) in p47phox and a C-terminal Src homology 3 (SH3) domain in p67phox . In the non-activated cell, the p47phox-p67phox complex fails to bind to flavocytochrome b558, due to an unusual autoinhibitory mechanism. In the resting cell, tandem SH3 domains in p47phox (referred to as the bis-SH3 domain) bind to an auto-inhibitory region (AIR), preventing the bis-SH3 domain from binding to the PRR of p22phox. Upon cell activation, serine residues of the AIR become phosphorylated, releasing the bis- SH3 domain so that it can now bind to p22phox [19–22]. In addition, p47phox possesses another membrane-binding region, the PX domain, which binds to the headgroups of phosphatidylinositols present in the membrane [23–25]. Together, these interactions with membrane lipids and p22phox promote the assembly of p47phox and p67phox with flavocytochrome b558. Concurrently, Rac-GTP translocates to the membrane where it interacts with the N-terminal tetratricopeptide repeat (TPR) region of p67phox [26–28]. An activation domain (AD) in p67phox, which is conformationally regulated by Rac binding , activates the hydride transfer from NADPH to FAD in Nox2. Subsequent electron transfer through two heme groups permits reduction of oxygen to form superoxide [30, 31]. p40phox binds through its Phox/Bem 1 (PB1) domain to a partner PB1 domain in p67phox and facilitates the assembly of p67phox-p47phox at the membrane through lipid binding via its PX domain [32–34]. Although no CGD patients have been described with mutations in p40phox, neutrophils from p40phox-knockout mice exhibit CGD-like severe defects in Nox2-derived oxidant-dependent bacterial killing , also suggesting that p40phox is a crucial component for Nox2 function.
The first homologue of gp91phox identified was Nox1, which is abundantly expressed in non-phagocyte cells including gastrointestinal epithelium and vascular smooth muscle [9, 36]. Subsequently, additional homologues were identified, and the Nox/Duox family in humans now consists of seven members: Nox1 though Nox5, Duox1 and Duox2 [9, 37]. All members of Nox/Duox family contain the six transmembrane α-helical heme-binding domain and an FAD domain containing an NADPH binding site, which together constitute the "Nox domain". Nox5 and Duox1/2 contain a calcium-binding EF-hand motif. The evolution of these Noxes and their calcium-binding domains were considered in the recent article  and are not discussed here. The remaining Noxes, Nox1-4, require p22phox for activity, and Nox1-3 all require additional Phox-like regulatory subunits, reviewed recently [10, 11, 37].
Nox1, like gp91phox, is activated by regulatory subunits: NOXO1 (Nox-organizer protein 1) is a homologue of p47phox and NOXA1 (Nox-activator protein 1) is a homologue of p67phox [38–41]. Recent studies have also demonstrated activation of the Nox1 system by Rac1 [42–45]. NOXO1 and NOXA1 are co-expressed with Nox1 in colon epithelium and gastric mucosa [39, 43, 46], consistent with in vivo regulation of Nox1 by these novel subunits. Activation of Nox1 by regulatory subunits differs in two major respects compared to gp91phox. First, NOXO1 lacks the AIR and its associated regulatory phosphorylation sites that are present in p47phox. Consistent with the domain structure, NOXO1 co-localizes with Nox1/p22phox complex constitutively at/near the plasma membrane in vivo [40, 41]. On the other hand, a recent study revealed that the bis-SH3 domain of NOXO1 bind to the extremely C-terminal PRR of NOXO1 . Phosphorylation-independent disruption of the intramolecular interaction facilitates NOXO1 binding to the PRR of p22phox. Second, NOXA1 lacks critical basic amino acid residues in its PB1 domain that are important for binding to p40phox. NOXA1 does not interact with p40phox, making human Nox1 activity independent of p40phox . Furthermore, human NOXA1 lacks the central SH3 domain that is present in p67phox [38–40], but whose function is unknown. Like Nox1, Nox3 is also regulated by p22phox and NOXO1, but in humans does not require NOXA1 [48–51]. In comparison, Nox4 requires p22phox for activity, but other known regulatory subunits are not needed [52–54].
Another subfamily of the Noxes is the NoxA/NoxB group, found in a slime mold amoeba and fungi. These fungal Nox genes have been identified in most ascomycetes except for hemiascomycete yeasts, and also in higher basidiomycetes and chytridiomycotes (reviewed in ). No Nox gene has been identified in fungi that belong to Zygomycota and Micropordia. These NoxA/NoxB genes are closely related to the subunit-regulated Noxes according to taxonomy . A non-animal homologue of p67phox/NOXA1 was described in slime mold amoeba Dictyostelium discoideum (D. discoideum) . The p67phox homolog (referred to here as the Dd-p67-like) of D. discoideum possesses the N-terminal TPR region and PB1 domain characteristic of p67phox, but lacks the SH3 domain . A p67phox homolog Nox-regulator (NOXR) was also reported in some fungi including ascomycetes and higher basidiomycetes [55, 57]. The basic motif structure of NOXR is similar to that of D. discoideum. A NOXR-deficient mutant of the ascomycete Epichlöe festucae shows decreased ROS-generation . Taken together, these data suggest that NOXR regulates NoxA and/or NoxB in the slime mold and fungi, although this has not been tested directly.
Herein, we assembled and analyzed experimentally cloned and computationally predicted amino acid sequences of the Phox-related regulatory subunits p47phox, NOXO1, p67phox, NOXA1, p40phox, p22phox, and NOXR from 10 vertebrates, one urochordate, one echinoderm, one mollusc, one cnidarian, one choanoflagellate, 13 fungi, and one slime mold amoeba. Using this large set of amino acid sequences, we report: (i) the occurrence and origin of Phox-like subunits during evolution, (ii) identification of putative key amino acid residues conserved in regulatory subunits, (iii) the description of putative new modular regions that are conserved among Phox-related regulatory subunits, (iv) the molecular evolution of characteristic domains of the Phox-related regulatory subunits, and (v) a prediction of novel fungal and amoebal Nox organizer/enhancer subunits based on sequence analysis.
Results and discussion
Occurrence of Phox-related regulatory subunits
The earlier chordate Ciona intestinalis (C. intestinalis) possesses a Nox2 ortholog, but not Nox1 or Nox3 [1, 61]. The sea urchin Strongylocentrotus purpuratus (S. purpuratus) belongs to Echinodermata, which diverged early from an ancestor common to urochordates and vertebrates . S. purpuratus, possesses two Nox2 genes, Nox2A and Nox2B, and an analysis of the taxonomy Noxes demonstrates that these Nox2 genes are related to the common ancestor gene of vertebrate Nox1, Nox2 and Nox3 . C. intestinalis possesses single copies of p47phox-, p67phox-, and p22phox-like genes , which presumably regulate its Nox2 ortholog (Figure 1A). The S. purpuratus genome possesses single copies of p47phox- and p67phox-like genes (Figure 1A). No p22phox ortholog of S. purpuratus was identified using a BLAST search of recently published genome sequences . Because the S. purpuratus p47phox-like protein has a highly conserved tandem SH3 region that in other species binds to the C-terminal proline-rich region (PRR) of p22phox, it seems likely that S. purpuratus has an as-yet unidentified p22phox homologue.
Using the estimated time of evolution of deuterostomes based on Hox gene clusters to construct a phylogenetic tree , we illustrate the occurrence during evolution of the Phox-related regulatory subunits(Figure 1B). The phylogenic tree suggests an evolutionary model in which duplication of both the p47phox gene and the p67phox gene occurred at a time corresponding to the appearance of vertebrates, with the NOXO1 and NOXA1 genes subsequently evolving as distinct regulatory subunits in vertebrates (Figure 1B). This appearance of NOXO1 and NOXA1 coincided with the emergence of Nox1 . The pufferfish T. rubripes and T. nigroviridis are considered to have evolved from an O. latipes ancestor approximately 186 million years ago . This tree suggests that NOXA1 and NOXO1 of T. rubripes and NOXA1 of T. nigroviridis have been lost during the decrease in genome sizes in these species (Figure 1B). In contrast to the co-appearance of p47phox and p67phox orthologs with the earliest Nox2 ortholog in S. purpuratus, p40phox appeared later, in vertebrates, and was not found in the genomes of either C. intestinalis or S. purpuratus (Figure 1B). The situation of the regulatory subunits of other primitive animals and a choanoflagellate is described below.
A homologue of p67phox/NOXA1 occurs in the slime mold amoeba (D. discoideum)  (Figure 1A). Fungal genomes of Aspergillus nidulans (A. nidulans), Magnaporthe grisea (M. grisea), and Fusarium graminearum (F. graminearum), all members of Ascomycota, also possess NOXR, a homolog of p67phox  (Figure 1A). However, Saccharomyces cerevisiae (S. cerevisiae) that also belongs to Ascomycota does not possess any Nox or NOXR orthologs [1, 55] (Figure 1A). Some basidiomycetes [Coprinopsis cinerea (C. cinerea), Laccaria bicolor (L. bicolor), and Postia placenta (P. placenta)] also possess NOXR and NoxA/B genes (Figure 1A). The situation of the regulatory subunits of other fungi is described below. Interestingly, the fungi seem to lack precise homologues of p47phox/NOXO1 or its binding partner p22phox (Figure 1A). Recently, the possibility of p22phox-like gene in the D. discoideum genome was suggested , but it is not clear whether the gene represents an ancestral p22phox, a later adaptation or a functionally unrelated protein. We discuss the possibilities below based on a comparison of the amino acid sequences of amoebal p22phox and other p22phox orthologs. In addition, we identified other predicted proteins encoded in fungal and amoebal genomes that may function as (a) binding partner(s) of the Nox activator proteins (described below), suggesting the early evolutionary origin not only of the activator protein, but also possibly of a rudimentary organizer subunit.
Synteny of NOXO1 ad NOXA1 genes in vertebrates
The synteny of genetic markers surrounding NOXA1 was highly conserved in H. sapiens, C. familliaris, M. musculus, G. gallus (Figure 2B). Both NOXA1 and the linked markers of R. norvegicus were present in chromosome 3, with a long insert between NOXA1and ENTPD8 (Figure 2B). In the genome of X. tropicalis, synteny was more varied in that NOXA1 and ENTPD8 were present in the same scaffold, but other markers were not seen. Synteny of fish NOXA1 was preserved among the class but different from other vertebrates. O. latipes and D. rerio NOXA1 genes are preceded by MAN1B1 and DPP7 on chromosome 9 and 5, respectively (Figure 2B). T. nigroviridis possesses two markers MAN1B1 and DPP7, but lacks the NOXA1 ortholog. Synteny of T. rubripes is similar to that of D. rerio, but a NOXA1 ortholog is absent. Thus, the synteny of NOXA1 had considerable variation, and this analysis indicates that NOXA1 orthologs were lost during the evolution of the two pufferfish genomes.
Identification of putative critical amino acids conserved among p47phox/NOXO1 orthologs
While the PX and bis-SH3 domains are shared among all p47phox and NOXO1 orthologs including those of S. purpuratus and C. intestinalis, the molecular taxonomy of the regions (excluding the variable C-terminal regulator docking regions) reveals that vertebrate p47phox and NOXO1 families form distinct subgroups (Figure 3B), likely accounting, for example, for differences in lipid binding specificities [24, 41]. The phylogenic tree supports an evolutionary model in which the p47phox-like proteins of S. purpuratus and C. intestinalis (#21 and 20 of Figure 3B, respectively) branch from a root that is closely related to the primordial ancestors of both p47phox (#1–10 of Figure 3B) and NOXO1 (#11–19 of Figure 3B).
To maintain p47phox strictly in an inactive state that cannot bind to p22phox, human p47phox masks its bis-SH3 domain with the AIR region [21, 68, 69], and this region is absent in human NOXO1 [38–41]. Therefore Nox2 activity is considered more tightly controlled than Nox1 activity. Most residues essential for the function of the AIR (e.g., Pro-299, Pro-300, Arg-301, Arg-302, Ser-303, Ser- 304 and Ser-328 of human p47phox), are well conserved among all vertebrate p47phox proteins (black boxes of Figure 3C). Interestingly C. intestinalis and S. purpuratus p47phox proteins possessed the AIR-like region (Figure 3C). S. purpuratus possesses two Nox2-like proteins Nox2A and 2B that are co-orthologs of vertebrate Noxes 1–3; the presence of a p47phox ortholog harboring the AIR suggests that this primitive Nox system functions analogously to the mammalian Nox2 system. All NOXO1 orthologs lack the AIR, with one exception, the teleost fish O. latipes, which retains the predicted AIR in both the p47phox and the NOXO1 genes (Figure 3C). Thus, this analysis supports an evolutionary model in which the NOXO1 "AIR-independent" Nox1 and Nox3 activation systems originated from the p47phox-like "AIR-dependent" Nox2 regulatory system. The O. latipes NOXO1, which contains the AIR, represents an evolutionary remnant reflecting the transition from AIR-dependence to -independence.
The heterodimerization of human p47phox and p67phox is dependent on the interaction between the C-terminal PRR of p47phox and the C-terminal SH3 domain of p67phox [40, 70–72]. Both of these regions were conserved in vertebrate p47phox and p67phox orthologs, but not in orthologs of lower organisms (Figure 3A). The alignment of vertebrate p47phox and NOXO1 amino acid sequences demonstrates that in addition to the PRR, the C-terminal region immediately following the PRR ("C-tail" in Figure 3D) was also highly conserved. The PRR of p47phox isologs possesses four strictly conserved residues (Pro-363, Pro-366, Arg-368, Pro-369; human-p47phox numbering), and the C-tail region had an additional four identical amino acids (Ile-374, Arg-377, Cys-378, and Thr-382; human-p47phox numbering) (Figure 3D). Consistent with these identities, the essential binding roles of three residues (Pro-363, Pro-366, Arg-368, indicated by black boxes of Figure 3D) were previously demonstrated by mutational analyses . The C-tail region is thought to enhance the interaction between the PRR of p47phox and the SH3 domain of p67phox , since simultaneous mutation of six of these residues (Leu-373, Ile-374, Arg-377, Thr-382, Lys-383, and Lys-385; human-p47phox numbering, indicated by black boxes of Figure 3D) significantly decreased the binding of the p47phox PRR to the p67phox SH3 domain [18, 72]. Four of these residues (Ile-374, Arg-377, Thr-382, and Lys-383) are highly conserved among both p47phox and NOXO1 proteins, but two other residues (Leu-373 and Lys-385; human-p47phox numbering) were conserved only in p47phox orthologs (Figure 3D). This suggests that the binding affinities of p47phox and NOXO1 toward their respective activator protein SH3 domains may differ due to differences in these residues. Ser-379 in the C-tail region of human p47phox is a phosphorylation site ; the phosphorylation of which negatively regulates binding of the PRR and C-tail regions to the p67phox SH3 domain . The threonine or serine residue is conserved at the position in all vertebrate p47phox and G. gallus, D. rerio and O. latipes NOXO1 (indicated by an arrow head in Figure 3D), indicating that NOXO1 proteins of these species retain this feature of p47phox which is subsequently lost in higher vertebrate NOXO1.
T. rubripes C17orf39 ortholog as a candidate NOXO1-like protein
Identification of putative critical amino acids conserved among p67phox/NOXA1 orthologs
In contrast to the C-terminal SH3 domain (SH3-B) of activator proteins which is seen in all vertebrate activator proteins, the central SH3 domain (SH3-A) was seen only in vertebrate p67phox (Figure 5A). Comparison of sequences of all vertebrate p67phox orthologs identified 31 highly conserved amino acids in this SH3 domain (Figure 5A and Additional file 3). In addition, a region between AD and SH3-A (termed here the ADSIS region for " AD-S H3 I ntervening S equence") was strictly conserved among vertebrate p67phox isologs, but was not seen in others, such as NOXA1 and NOXR. A consensus sequence was identified in this region (Figure 5D). A PRR-like region (residues 227–231 of human p67phox) was seen in ADSIS region, but it does not conserve in all vertebrate p67phox isologs based on the deduced sequences (Figure 5D). The role of the putative region is currently unknown, but may involve a function related to SH3-A because it coincides with the presence of SH3-A. Despite the high conservation of SH3-A among vertebrate p67phox orthologs, the role of this domain is not defined [10, 75].
The PB1 domain was present in all vertebrate p67phox, in vertebrate NOXA1 and in fungal NOXR and amoebal p67-like protein, but not in C. intestinalis and S. purpuratus p67phox-like proteins (Figure 5A). The crystal structure of the heterodimer between the PB1 domains of p67phox and p40phox domain demonstrated that Lys-355 and Lys-382 in human p67phox are essential to bind the PB1 domain of p40phox [32, 76]. Residues corresponding to Lys-355 and Lys-382 of human p67phox are conserved not only in the vertebrate p67phox, but also in the NOXA1 orthologs of chicken (G. gallus) and fish (D. rerio and O. latipes) (Figure 5E). This suggests that in addition to its well-described role in activating Nox2, p40phox probably also participates in activating Nox1 in chicken and fish (Figure 5E). On the other hand, NOXA1 of X. tropicalis has lost both of these p40phox-interacting residues. Thus, these data support an evolutionary model in which p40phox originally participated in regulating both Nox1 and Nox2 in vertebrates, but after the appearance of mammals, Nox1 (but not Nox2) became independent of p40phox. The lack of a PB1 domain in both C. intestinalis and S. purpuratus p67phox-like proteins is consistent with the absence of p40phox orthologs in these species (Figure 1B).
The analysis of PB1 domain sequences in Nox activators also suggests an unexpected relationship between the AD and the PB1 domain. AD regions of all vertebrate p67phox contain a positively charged residue, e.g., Lys-196 of h-p67phox (indicated by gray boxes in Figure 5C). This basic residue is also completely conserved in NOXA1 orthologs of G. gallus, D. rerio and O. latipes (Figure 5C), which retain functional PB1 domains (Figure 5E), but not in the other NOXA1 and p67phox-like proteins (Figure 5C) (with the single exception of the C-terminal AD-like region of S. purpuratus p67phox-like protein). Interestingly, the fungal NOXR, which possess these key basic residues in their PB1 domains (Figure 5E), also conserves this basic residue in their AD regions (Figure 5C). Thus, the presence of a functional PB1 domain correlates strongly with the presence of a positively charged residue corresponding to position 196 in the human p67phox sequence, and may point to the functional cooperation of these two regions, e.g. in binding to p40phox or other functionally analogous proteins in fungi (Figure 5F). Alternatively, the positively charged residue in AD region might be related to the tail-to-tail interaction between vertebrate p47phox and p67phox (and NOXO1-NOXA1 of early vertebrates) because a sequence analysis of Nox organizers that is shown in Figure 3D suggests that the conserved residue also co-occurs with the phosphorylation site in the C-tail region of all vertebrate p47phox and early vertebrate NOXO1. Thus, co-appearance and co-disappearance of evolutionarily conserved residue provides a hint to find an unknown relationship between regulatory-subunits.
SH3-B was seen in all vertebrate p67phox and NOXA1 orthologs and in S. purpuratus p67phox-like protein (Figure 5A). Among these orthologs, seven conserved amino acid residues in SH3-B were identified (Figure 5A), and the residues include a tryptophan (Trp-494; human p67phox- numbering) that was previously shown to be crucial for interaction with the C-terminal PRR of Nox organizer proteins .
Identification of putative critical amino acids conserved among p40phox orthologs
To the best of our knowledge, a role for the SH3 domain of p40phox has not been defined . Eighteen amino acids are conserved in the SH3 domain (see Additional file 3). Between the PX and SH3 domains, we identified a new conserved region of 15 amino acids that is rich in proline, arginine and lysine, and is therefore designated the "proline-basic (P-basic) domain" (Figure 6A). This putative region is not essential for PtdIns(3)P-binding by the p40phox PX domain [25, 80]. An intramolecular interaction between the PX and PB1 domains seem not to be related to P-basic region according to the structure of full length p40phox [81, 82]. The structural analysis demonstrates that P-basic region forms a separate motif and is situated closely to the PX domain , indicating that it might support the cell membrane-binding function of p40phox together with the PX-domain. Alternatively, the P-basic region might interact with the SH3 domain of p40phox in a manner similar to the interaction between the AIR and the tandem SH3 of human p47phox. Interestingly, Thr-154 of human p40phox, located in the center of P-basic region, has been identified as a phosphorylation site , and this residue is entirely conserved among all vertebrate p40phox orthologs (Figure 6A).
The PB1 domain of human p40phox harbors acidic amino acids forming the "OPCA (O PR/PC/A ID) motif". An OPCA motif contains four distinctive acidic residues that are essential for binding to conserved basic residues in the p67phox PB1 domain . The four acidic residues, Asp-289, Glu-291, Asp-293 and Asp-302 (human p40phox numbering) are conserved in all orthologs with the exception of a glutamine rather than a glutamic acid in the position of C. familliaris p40phox that corresponds to Glu-291 of human p40phox (Figure 6A).
Identification of putative critical amino acids conserved among p22phox orthologs
As shown in Figure 1, a p22phox-like protein was not identified in S. purpuratus, but because the genome sequence of this organism is incomplete and because a p47phox-like protein with a conserved bis-SH3 domain is present, we suggest that this organism probably possesses an undiscovered p22phox-like protein, or a protein that serves an analogous function. Our analysis did not provide convincing evidence for a p22phox isolog in slime mold amoeba. Lardy et al. argued that this organism may possess a membrane-associated p22phox- like protein (GenBank™ accession number AY221170); predicted p22phox-like gene-null D. discoideum mutant is unable to produce spores, similar to nox-null mutants . However according to our analysis, this protein is highly divergent in amino acid sequence compared with the consensus sequence obtained from all other organisms, showing disagreement in 24 out of the 38 conserved residues shown in Figure 7A. Thus, further investigation is needed to judge whether the gene emerged as a later adaptation or whether it represents a primitive p22phox gene.
Molecular evolution of domains of Nox organizers and activators in Deuterostomia
Like vertebrate isologs of p47phox, the p47phox-like proteins of C. intestinalis and S. purpuratus possess the PX domain, bis- SH3 domain, and AIR. However, these isologs differ considerably in the C-terminal regions that, in higher forms, interact with p67phox isologs. The C-terminus of the C. intestinalis p47phox lacks the PRR, but motif scanning by PROSITE  demonstrates the p47phox-like protein contains, instead, additional SH3 domain [residues 340–399 of Ci-p47phox protein, GenBank™ No. NM_001033828]. The S. purpuratus p47phox-like protein also lacked the C-terminal PRR, but instead had two additional SH3 domains (residues 367–430 and 561–618 of Sp-p47phox, GenBank™ No. XP_001183696) separated by two PRRs (residues 453–460 and 476–484 of Sp-p47phox) (bottom two left-hand panels in Figure 8).
All vertebrate orthologs of p67phox have the same domains seen in human p67phox (Figure 8, left panels). Importantly, all p67phox and NOXA1 proteins possess the AD region, which has been shown to be critical for activating superoxide generation in Nox2 and Nox1 [30, 31]. The TPR motif is present in all species, and the two critical residues for Rac-binding, Asp-67 and Arg-102 of human p67phox , are conserved among all vertebrate isologs of p67phox and NOXA1 (top four panels in Figure 8), supporting the idea that binding to Rac is a conserved feature of vertebrate p67phox andNOXA1. The N-terminal domain structures of C. intestinalis and S. purpuratus p67phox-like proteins are similar to those of vertebrate p67phox and NOXA1 (Figure 8). Both p67phox-like proteins possess a TPR domain retaining the two critical Rac-binding residues (Figure 8). In addition, S. purpuratus p67phox-like protein possesses a second set of TPR and AD-like domains. Both TRP domains retain conserved essential Rac-binding residues, but the second AD-like domain was not well conserved as shown in Figure 5C. Thus, Rac binding is likely to be an evolutionarily conserved feature among all p67phox/NOXA1 proteins.
In mammals, binding between the PB1 domains of p67phox and p40phox has been shown to mediate heterodimer formation. The crystal structure of the heterodimer of the PB1 domains from p67phox and p40phox  demonstrated that Lys-355 and Lys-382 in human p67phox are essential to bind to the PB1 domain of p40phox, and mutation of Lys-355 of human p67phox abolished binding to p40phox . These crucial residues are entirely conserved in the PB1 domains of all vertebrate p67phox proteins, which imply that they all bind to p40phox (top four left-hand panels in Figure 8). In contrast to vertebrate p67phox, mammalian NOXA1 does not retain these two key amino acids (a top right-hand panel of Figure 8), explaining the failure of NOXA1 to bind to p40phox . However, the chicken G. gallus, and the fish D. rerio and O. latipes NOXA1 completely conserve these two residues, and an OPCA motif of p40phox (Figure 6), which is essential for binding to the p67phox PB1 domain are highly conserved. The conservations of key residues suggest that in these fish, binding to p40phox may be retained (from the second to fourth panels in right hand of Figure 8). Thus, the loss of function of p40phox can be traced in the evolution of NOXA1. Both C. intestinalis and S. purpuratus p67phox-like proteins lack a PB1 domain, consistent with the absence of p40phox orthologs.
The PX-domain of human p40phox specifically binds to PtdIns(3)P [23, 24, 79, 80]. Therefore, p40phox accumulates during phagocytosis at early endosomes, which are rich in PtdIns(3)P [35, 81, 88]. T. rubripes p40phox is expressed in a wide range of tissues including the gastrointestinal tissues , while Nox2 expression is restricted to the blood and kidney. Although Nox1 expression in fish or the other non-mammal organisms is not clear, our analysis suggest that Nox1 of fish and birds might function, along with the p40phox orthologs, in endocytosis, e.g., in receptor-internalization. This novel relationship between Nox1 and p40phox may provide a clue to understanding the physiological role of Nox1 in vertebrates.
The vertebrate p67phox and NOXA1 orthologs all have the C-terminal SH3 domain that, in human, mediates binding to the PRR of p47phox and NOXO1, respectively. This supports the idea that all of the vertebrate forms of these proteins mediate protein interactions via tail-to-tail binding between the SH3 domain of the activator protein and the PRR of the organizer protein. We further suggest that even though the structures of the C-termini of p47phox- and p67phox-like proteins in C. intestinalis and in S. purpuratus differ from the vertebrate proteins, the same sort of tail-to-tail binding of these p67phox-like proteins to the corresponding p47phox-like proteins occurs as diagrammed at the bottom of Figure 8. For example, we suggest that the C-terminal putative PRRs of C. intestinalis p67phox-like proteins (residues 256–348 of Ci-p67phox protein, GenBank™ No. NM_001033827) binds to the C-terminal SH3 domain of the p47phox-like protein as indicated by a broken line in Figure 8, reversing the mode of interaction seen in the vertebrate proteins but accomplishing the same function. Similarly, SH3 domains and putative PRRs in the C-terminus of S. purpuratus p47phox-like protein may interact with the PRRs and the SH3 domains in the C-terminus of S. purpura tus p67phox, respectively. Thus, while domains in the C-termini of p47phox and p67phox orthologs are not conserved during evolution, the model shown in Figure 8 suggests that the "docking function" has been retained. The model suggests three functional modules within the organizer protein/activator protein complex: 1), a "membrane-binding/cytochrome docking region" on the organizer protein that is comprised of the phospholipids-binding PX domain and the tandem SH3 region that binds to the PRR of p22phox; 2) a "Nox-activating region" that contains the Rac-binding TPR domain and the AD, which together cooperate to stimulate the activity of the Nox catalytic domain; and 3) a "regulator-docking region" which uses evolutionarily variable but paired docking modules that link the "membrane-binding/cytochrome docking region" of p47phox to the "Nox-activating region" of p67phox (bottom panel in Figure 8).
Occurrence and origin of Phox-like regulatory subunits during evolution of Metazoa and Choanoflagellata
Before the emergence of bilateral animals, cnidarians split from the main animal lineage and the genome retained many of the features present in last common ancestor of animals . The starlet sea anemone N. vectensis that belongs to the Cnidaria, conserves p47phox-, p22phox-like genes, and two ancestral Nox2 genes, Nox2A and 2B (Figure 9A). The characteristic motifs of the putative regulatory subunits are predicted by the Pfam program and shown in Figure 9B (alignments are shown in Additional file 8). N. vectensis p47phox possesses PX and bis-SH3 domains. The isolog also possesses a putative polybasic region within the sequence corresponding to the AIR of the human p47phox gene. A polybasic region is a class III consensus motif for binding to a SH3 domain ; therefore, this may implicate the polybasic region as a prototype of the AIR. Although the current version of the N. vectensis genome does not encode an obvious p67phox-like gene, the presence of multiple SH3 domains in N. vectensis p47phox points to an unidentified p67phox ortholog or an unknown PRR-containing binding partner. Although cnidarians have no specialized immune cells, the N. vectensis genome possesses orthologous genes to Toll/Toll-like receptors, complement C3-like molecules, and a redox-sensitive transcription factor nuclear factor-κB that are orthologs of key components of innate immunity in vertebrates . Further study needs to identify an original role of Phox-type NADPH oxidase in the primitive multicellular animals.
One clear implication of these genomic and detailed sequence analyses described above is that the hypothetical ancestor of primitive animals might conserve the Phox-type NADPH-oxidase system. In the middle of the nineteenth century, a striking structural resemblance between the collar cells, so-called "choanocytes", of primitive animal sponges and a group of protists the choanoflagellates has been noted (, reviewed in ). A line of recent phylogenic analyses suggests that the metazoa including sponges, cnidarians, and other animals constitutes a monophyletic clade closely related to the Choanoflagellata [90, 95, 96]; therefore, the choanoflagellates are considered to be the organisms that are close in evolution to the all modern animals. The unicellular marine choanoflagellate M. brevicollis has been reported as a closer relatives to the hypothetical ancestor of all multicellular animals based on the comparative analyses of animal-specific genes [97, 98]. Interestingly, the genome of M. brevicollis possesses p47phox- and p22phox-like genes (Figures 9A,B), but a BLAST searching in the current M. brevicollis genomic database failed to identify a dinstinct p67phox ortholog. Furthermore, Nox2-like gene is present in the M. brevicollis genome sequence (Figure 9A). Interestingly, a taxonomy of Nox domains demonstrates that M. brevicollis Nox2-like gene diverges from the monophyletic clade of all metazoan Nox2 genes containing other ancestral Nox2 and all vertebrate Nox1-3, but it differs from Nox4 and fungal NoxA/NoxB subfamilies (see Additional file 6). The M. brevicollis p47phox-like protein possesses AIR-like region that conserves a serine residue consistent with a consensus motif of a putative substrate of protein kinase C (Figure 9C); therefore, this implies that ROS-generating activity of the most primitive Nox2-like protein is also strictly regulated by phosphorylation as seen in mammalian phagocyte NADPH-oxidase. The physiological function of the primordial Nox2 enzyme in M. brevicollis is unknown, but the early appearance of the Phox-like Nox system in the unicellular flagellate suggests the presence of a fundamental and essential role of the regulatory subunit-dependent Nox in a unicellular living.
Figure 9A also summarizes the occurrence of Nox4, Nox5, and Duox isologs in genomes of the metazoa and the choanoflagellate, adding to information published previously . Duox orthologs are present in all analyzed genomes of the Bilateria (Figure 9A). Nox5 genes occur broadly in the genomes of Metazoa, but not in Nematoda. Nox4 genes appear after the emergence of chordates, but is not seen in the echinoderm S. purpuratus  (Figure 1A). Interestingly, gene search that covers a wide range demonstrates that Nox4-like genes are also present in a lophotrochozoan (L. gigantea), a cnidarian (N. vectensis), but not in the ecdysozoans (see Figure 9A, and the taxonomy of Nox is shown in Additional file 6). The choanoflagellate M. brevicolli do not possess Nox4, Nox5, and Duox orthologs, at least based on the current genomic database. In the kingdom Plantae, the moss Physcomitrella patens (P. patens) possess double EF-hand- containing Noxes like the higher land plants Arabidopsis thaliana (A. thaliana) and Oryza sativa (O. sativa) (Figure 9A). Despite a kingdom distinct from plants, the oomycete Phytophthora sojae (P. sojae) also possesses single or double EF-hand(s)-containing Noxes [55, 99], at least three genes (Figure 9A). Here we referred to them as P. patens and P. sojae rboh (respiratory burst oxidase homolog) to represent the remarkable resemblance to the higher plant Noxes, so-called rboh, in the respects of taxonomy of the Nox domains (Additional file 6). In the kingdoms Plantae and Chomista, a distinct regulatory subunit was not seen. However, higher plant genomes possess a novel protein family conserved domain structures similar to p67phox as descried below. Naegleria gruberi (N. gruberi) is a widespread soil and freshwater amoeboflagellate that belongs to the phylum Perocolozoa, the kingdom Protozoa . Although N. gruberi is a free-living organism, it is not related to species of the phylum Amoebozoa with sequenced genomes, such as D. discoideum and Entamoeba histolytica, but it is close to pathogenic relatives (e.g. Naegleria fowleri) that can cause amoebic meningitis. We identified single Nox gene of N. gruberi from the genome (Figure 9A), together with five putative Fre ortholog genes that have similar domain structures to that of Nox domain (see Additional file 9). According to a phylogenetic taxonomy of Nox domain, the N. gruberi Nox is close to D. discoideum NoxA/NoxB proteins (see Additional file 6); therefore, it is referred to as N. gruberi NoxA (Figure 9A). Nox regulatory subunit gene has not been found in the current version of genomic database, but it might contain a prototype of regulatory subunits according to the Nox taxonomy.
SH3PXD2 as a p47phox family member
What is the regulatory subunit binding partner for fungal NOXR?
The "SH3-SH3-PX-SRR-PB1" protein contains a similar domain structure to yeast S. cerevisiae Bem1p (Bud Emergence 1, GenBank™ accession number NP_009759), but differs in that the PB1 domain of S. cerevisiae Bem1p does not contain the OPCA motif . A speculative model for the regulation of fungal Nox by the Bem1 ortholog is shown in Figure 11D, wherein Bem1 ortholog is proposed to play a role as an organizer protein, analogous to the roles of p47phox and p40phox in mammals. According to this model, PB1-PB1 interaction mediates the linkage between the "regulator-docking regions" suggested in Figure 8. PX-domain of the S. cerevisiae Bem1 facilitates a protein to anchor to a membrane lipid layer . Fungi do not have a PRR-containing p22phox ortholog (see Figure 1A), but NoxA and NoxB in the three fungi has a distinctive highly conserved a class II PRR motif  (e.g. 423–428 residues of Mg-NoxA) and class III (e.g. 447–452 residues of Mg-NoxB) near the predicted NADPH-binding sub-regions . Thus the hypothetical functions of the bis- SH3 and PX domains indicate the analogous function as a "membrane-binding/cytochrome docking region" of Nox organizer. Because S. cerevisiae Bem1 is involved in establishment of cell polarity , the proposed interaction between fungal NOXR and Bem1 orthologs may point to a function of Nox-derived ROS in cell polarity.
A second model is suggested by the "CDC24-RhoGEF-PB1" protein, which is an ortholog of S. cerevisiae Cdc24p (GenBank™ accession number AAA82871). According to this "exchanger model", the PB1-mediated heterodimer formation localizes the RhoGEF domain which then catalyzes the conversion of Rac-GDP to the active Rac-GTP (Figure 11E). The model is supported by a recent report showed that fungal NOXR binds to the fungal Rac homologue in the GTP form .
An "energy sensor" function for NOXR is shown in Figure 11F, and is based on the occurrence of the CBS (cystathione beta-synthase) domain in the putative binding partner. The CBS domain senses cellular energy status by binding adenosine nucleotides; it occurs in a range of proteins including AMP-activated protein kinase, which is activated by AMP and inhibited by ATP via the CBS domain [106, 107]. Such a function might be related to the requirement of Nox enzyme for reduced pyridine nucleotide, thus affecting cellular energetics.
Predicted binding partners for D. discoideum p67-like protein
Do p67phox-like genes exist in plants?
The land plants, A. thaliana and O. sativa possess the multiple EF-hand-containing Nox proteins, At-rboh-A to -J [1, 112, 113] and Os-rboh-A  to -H, respectively. However, there is no report of Phox-related regulatory subunit homologs in plant genomes. Unexpectedly BLAST search using PB1 domain sequences demonstrates that A. thaliana possesses a unique family of genes encoding four proteins that possess both TPR and PB1 domains (GenBank accession No. NP_194935, NP_180101, NP_197536, NP_564794). The O. sativa genome also possesses a similar gene (GenBank accession No. BAD31284). Herein, we refer to this family as "hypothetical p67phox-like (hypo-p67-L)". The expressions of at least the two identified genes, At-hypo-p67-L1 and -L4, were confirmed according to the EST database .
In summary, we reported here an exhaustive analysis of the Phox-related regulatory subunits for Nox enzymes from a total of 32 species in the Eukaryota including 10 vertebrates. The present study, viewed in the context of the evolution of Noxes , reveals that regulatory subunits co-evolved with their respective Noxes; Nox2 with p67phox/p47phox; Nox1 with NOXO1/NOXA1; ancestral Nox2 as co-ortholog of vertebrate Nox1-3 with p67phox/p47phox-like proteins; fungal and amoebal NoxA/B with NOXR (and possibly new components). By comparing amino acid sequences of the regulatory subunits, putative key amino acid residues and conserved region previously undefined were identified, e.g. ADSIS. These conserved residues are likely to be important in conserved catalytic or regulatory functions of Nox proteins, although, the detailed biochemical roles of most of these residues are not yet fully understood. The analysis of the Phox-related regulatory subunits points to a novel evolutionary history of Nox regulatory proteins. The earliest Nox2 and p47phox- and p22phox-like genes are appearing in the unicellular choanoflagellate before the emergence of the metazoa. Duplication of primordial p47phox- and p67phox-like genes occurred in vertebrates, resulting in the emergence of NOXO1-p47phox and NOXA1-p67phox genes, respectively. Supporting this evolutionary origin, the NOXO1 protein in the fish O. latipes still retains the p47phox-characteristic "AIR". A predicted motif structure of ancestral p47phox suggests a greatly expanded family of p47phox including SH3PXD2 in the animal lineages. Detailed sequence analyses proposed an evolution model that p40phox originally participated in regulating both Nox1 and Nox2 in early vertebrates, but in mammals the regulation of Nox1 became independent of p40phox. Thus, this report provides keys to understand the evolutionary history of Nox and the regulatory subunits, and provides new information that may be useful in elucidating new molecular details of the activation mechanisms of Nox enzymes including the human orthologs.
Note: While we were in the process of revising this manuscript, a review article about structure and function of the PB1 domain appeared . The review will help to understand the structural features of the PB1 domain that are often mentioned in the present study. In the article, authors also described the presence of the hypothetical genes coding TPR and PB1 domain-containing proteins in the A. thaliana genomes and argued about a possibility of A. thaliana p67phox.
Homologues/orthologs of Nox/Duox and regulatory subunits sequences were assembled from the following species: H. sapiens, (human), C. familliaris (dog), R. norvegicus (rat), M. musculus (mouse), G. gallus (chicken), X. tropicalis (frog), D. rerio (zebrafish), T. rubripes (fugu), T. nigroviridis (tetraodon), O. latipes (medaka), C. intestinalis (ascidian), S. purpuratus (sea urchin), D. pulex (waterflea), C.briggsae (nematode), C. elegans (nematode), N. vectensis (sea anemone), L. gigantea (gastropod snail), M. brevicollis (choanoflagellate), D. discoideum (slime mold amoeba), C. merolae (red alga), A. nidulans (fungus-An), M. grisea (fungus-Mg), F. graminearum (fungus-Fg), S. cerevisiae (fungus-Sc), C. albicans (fungus-Ca), Y. lipolytica (fungus-Yl), S. pombe (fungus-Sp), Coprinopsis cinerea (C. cinerea, fungus-Cc), L. bicolor (fungus-Lb), P. placenta (fungus-Pp), Puccinia graminis (P. graminis, fungus-Pg), P. blakesleeanus (fungus-Pb), B. dendrobatidis (fungus-Bd), P. sojae (oomycete), Phaeodactylum tricornutum (P. tricornutum, diatom), Physcomitrella patens (P. patens moss), A. thaliana (land plant), and O. sativa (land plant). Using NCBI-HomoloGene , existing homologues/orthologs of regulatory subunits were assembled: p47phox, NOXO1, p67phox, NOXA1, p40phox and p22phox orthologs of human, dog, mouse, rat, and p47phox and p67phox and NOXO1 orthologs of chicken. In addition, we assembled amino acid sequences that have been cloned and reported: p47phox, p67phox, and p22phox of T. rubripes  and C. intestinalis ; p67-like protein of D. discoideum ; NOXR of fungi including A. nidulans, M. grisea, F. graminearum [55, 57]. Based on these sequences, BLASTP searches were performed to obtain predicted regulatory subunits, related Nox genes (including genes that has been reported such as P. sojae rboh , C. merolae rboh1/2 , P. tricornutum rboh1 ), new regulator-binging partners, using the NCBI protein database for vertebrates, C. intestinalis, C. briggsae, D. discoideum, A. thaliana, O. sativa , S. purpuratus , and for fungi (A. nidulans, M. grisea, F. graminearum, S. cerevisiae, C. albicans, Y. lipolytica, S. pombe, C. cinerea) , the Doe Joint Genome Institute database for L. bicolor, P. placenta, D. pulex, N. vectensis, L. gigantea, M. brevicollis, P. blakesleeanus, P. sojae, P. tricornutum, P. patens , and the Broad Institute FGI database for P. graminis and B. dendrobatidis . Sequences that had more than 50% identity to the sequence of the closest template were chosen as orthologs or isologs. Assembled sequences, including newly defined, were annotated based on molecular taxonomy analysis of the domains characteristic in each regulator. Characteristic motifs of the regulatory subunits are predicted by the Pfam program . All amino acid sequences and the accession numbers of assembled and analyzed regulator orthologs are listed in Additional material (see Additional files 1, 5, 7, 9, 10, 12, 13, 15, 16, 18).
Phylogenic analysis and synteny
Multiple sequence alignment and phylogenetic analyses were carried out with ClustalW  and trees were reconstructed by the neighbor-joining method as previously described . Amino acid sequences of regulatory subunits were trimmed and aligned to the domains indicated in each figure. Bootstrap values of 1,000 replications are shown at the branches as percentages. Equivalent to 0.1 amino acid substitutions per site indicate evolutionary distances as inferior bars. To elucidate the synteny, we used the Ensembl AlignSliceView program . Conserved marker genes among the genomes of vertebrates are defined (see Additional file 2).
reactive oxygen species
Nox Organizer 1
Nox Activator 1
the tetratricopeptide repeats
Src homology 3
Phox homology domain
- OPCA motif:
AD-SH3 Intervening Sequence
SH3 and PX domains 2
Rho guanine nucleotide exchange factor
We express our appreciation to Mrs. Heather Jackson for reading and commenting on the manuscript and to Dr. Yasushi Okamura for suggesting sea urchin genome research. We thank Drs. Seiji Takeda and Takehiko Ueyama for commenting about A. thaliana p67phox-like genes and a conserved lysine residue in AD of p67phox, respectively. We also thank anonymous reviewers for suggestive comments to improve content of the manuscript. We are grateful to Mrs. Asami Kawahara for great cooperation and patience during long nights and weekends of endless computational analysis at home. This work was supported by NIH grants CA105116 and GM067717.
- Kawahara T, Quinn MT, Lambeth JD: Molecular evolution of the reactive oxygen-generating NADPH oxidase (Nox/Duox) family of enzymes. BMC Evolutionary Biology. 2007, 7 (109):
- Bedard K, Lardy B, Krause KH: NOX family NADPH oxidases: Not just in mammals. Biochimie. 2007, 89 (9): 1107-1112. 10.1016/j.biochi.2007.01.012.PubMedGoogle Scholar
- Vignais PV: The superoxide-generating NADPH oxidase; structural aspects and activation mechanism. Cellular and Molecular Life Sciences. 2002, 59 (9): 1428-1459. 10.1007/s00018-002-8520-9.PubMedGoogle Scholar
- Heyworth PG, Cross AR, Curnutte JT: Chronic granulomatous disease. Current Opinion in Immunology. 2003, 15 (5): 578-584. 10.1016/S0952-7915(03)00109-2.PubMedGoogle Scholar
- Lambeth JD: Nox enzymes, ROS, and chronic disease: an example of antagonistic pleiotropy. Free Radical Biology and Medicine. 2007, 43 (3): 332-347. 10.1016/j.freeradbiomed.2007.03.027.PubMed CentralPubMedGoogle Scholar
- Nauseef WM: Assembly of the phagocyte NADPH oxidase. Histochemistry and Cell Biology. 2004, 122 (4): 277-291. 10.1007/s00418-004-0679-8.PubMedGoogle Scholar
- Geiszt M: NADPH oxidases: new kids on the block. Cardiovascular Research. 2006, 71 (2): 289-299. 10.1016/j.cardiores.2006.05.004.PubMedGoogle Scholar
- Leto TL, Geiszt M: Role of Nox family NADPH oxidases in host defense. Antioxidants & Redox Signaling. 2006, 8 (9–10): 1549-1561.Google Scholar
- Lambeth JD: NOX enzymes and the biology of reactive oxygen. Nature Review Immunology. 2004, 4 (3): 181-189. 10.1038/nri1312.Google Scholar
- Sumimoto H, Miyano K, Takeya R: Molecular composition and regulation of the Nox family NAD(P)H oxidases. Biochemical and Biophysical Research Communications. 2005, 38 (1): 677-686. 10.1016/j.bbrc.2005.08.210.Google Scholar
- Lambeth JD, Kawahara T, Diebold B: Regulation of Nox and Duox enzymatic activity and expression. Free Radical Biology and Medicine. 2007, 43 (3): 319-331. 10.1016/j.freeradbiomed.2007.03.028.PubMed CentralPubMedGoogle Scholar
- Ishibashi F, Nunoi H, Endo F, Matsuda I, Kanegasaki S: Statistical and mutational analysis of chronic granulomatous disease in Japan with special reference to gp91-phox and p22-phox deficiency. Human Genetics. 2000, 106 (5): 473-481. 10.1007/s004390000288.PubMedGoogle Scholar
- Bionda C, Li XJ, van Bruggen R, Eppink M, Roos D, Morel F, Stasia MJ: Functional analysis of two-amino acid substitutions in gp91 phox in a patient with X-linked flavocytochrome b558-positive chronic granulomatous disease by means of transgenic PLB-985 cells. Human Genetics. 2004, 115 (5): 418-427. 10.1007/s00439-004-1173-z.PubMedGoogle Scholar
- Forbes L, Truong O, Wientjes FB, Moss SJ, Segal AW: The major phosphorylation site of the NADPH oxidase component p67phox is Thr233. Biochemical Journal. 1999, 338 (1): 99-105. 10.1042/0264-6021:3380099.PubMed CentralPubMedGoogle Scholar
- Regier DS, Greene DG, Sergeant S, Jesaitis AJ, McPhail LC: Phosphorylation of p22phox is mediated by phospholipase D-dependent and -independent mechanisms. Correlation of NADPH oxidase activity and p22phox phosphorylation. Journal of Biological Chemistry. 2000, 275 (37): 28406-28412. 10.1074/jbc.M004703200.PubMedGoogle Scholar
- Bouin AP, Grandvaux N, Vignais PV, Fuchs A: p40(phox) is phosphorylated on threonine 154 and serine 315 during activation of the phagocyte NADPH oxidase. Implication of a protein kinase c-type kinase in the phosphorylation process. Journal of Biological Chemistry. 1998, 273 (46): 30097-30103. 10.1074/jbc.273.46.30097.PubMedGoogle Scholar
- Knaus UG, Heyworth PG, Evans T, Curnutte JT, Bokoch GM: Regulation of phagocyte oxygen radical production by the GTP-binding protein Rac 2. Science. 1991, 254 (5037): 1512-1515. 10.1126/science.1660188.PubMedGoogle Scholar
- Kami K, Takeya R, Sumimoto H, Kohda D: Diverse recognition of non-PxxP peptide ligands by the SH3 domains from p67(phox), Grb2 and Pex13p. The EMBO Journal. 2002, 21 (16): 4268-4276. 10.1093/emboj/cdf428.PubMed CentralPubMedGoogle Scholar
- El Benna J, Faust L, Babior B: The phosphorylation of the respiratory burst oxidase component p47phox during neutrophil activation. Journal of Biological Chemistry. 1994, 269: 23431-23436.PubMedGoogle Scholar
- Ago T, Nunoi H, Ito T, Sumimoto H: Mechanism for phosphorylation-induced activation of the phagocyte NADPH oxidase protein p47(phox). Triple replacement of serines 303, 304, and 328 with aspartates disrupts the SH3 domain-mediated intramolecular interaction in p47(phox), thereby activating the oxidase. Journal of Biological Chemistry. 1999, 274 (47): 33644-33653. 10.1074/jbc.274.47.33644.PubMedGoogle Scholar
- Groemping Y, Lapouge K, Smerdon SJ, Rittinger K: Molecular basis of phosphorylation-induced activation of the NADPH oxidase. Cell. 2003, 113 (3): 343-355. 10.1016/S0092-8674(03)00314-3.PubMedGoogle Scholar
- Rotrosen D, Leto TL: Phosphorylation of neutrophil 47-kDa cytosolic oxidase factor. Translocation to membrane is associated with distinct phosphorylation events. Journal of Biological Chemistry. 1990, 265 (32): 19910-19915.PubMedGoogle Scholar
- Ago T, Takeya R, Hiroaki H, Kuribayashi F, Ito T, Kohda D, Sumimoto H: The PX domain as a novel phosphoinositide- binding module. Biochemical & Biophysical Research Communications. 2001, 287 (3): 733-738. 10.1006/bbrc.2001.5629.Google Scholar
- Kanai F, Liu H, Field SJ, Akbary H, Matsuo T, Brown GE, Cantley LC, Yaffe MB: The PX domains of p47phox and p40phox bind to lipid products of PI(3)K. Nature Cell Biology. 2001, 3 (7): 675-678. 10.1038/35083070.PubMedGoogle Scholar
- Karathanassis D, Stahelin RV, Bravo J, Perisic O, Pacold CM, Cho W, Williams RL: Binding of the PX domain of p47(phox) to phosphatidylinositol 3,4-bisphosphate and phosphatidic acid is masked by an intramolecular interaction. The EMBO Journal. 2002, 21 (19): 5057-5068. 10.1093/emboj/cdf519.PubMed CentralPubMedGoogle Scholar
- Ligeti E, Tardif M, Vignais PV: Activation of O2- generating oxidase of bovine neutrophils in a cell-free system. Interaction of a cytosolic factor with the plasma membrane and control by G nucleotides. Biochemistry. 1989, 28: 7116-7123. 10.1021/bi00443a050.PubMedGoogle Scholar
- Koga H, Terasawa H, Nunoi H, Takeshige K, Inagaki F, Sumimoto H: Tetratricopeptide repeat (TPR) motifs of p67phox participate in interaction with the small GTPase Rac and activation of the phagocyte NADPH oxidase. Journal of Biological Chemistry. 1999, 274: 25051-25060. 10.1074/jbc.274.35.25051.PubMedGoogle Scholar
- Lapouge K, Smith SJ, Walker PA, Gamblin SJ, Smerdon SJ, Rittinger K: Structure of the TPR domain of p67phox in complex with Rac.GTP. Molecular Cell. 2000, 6 (4): 899-907.PubMedGoogle Scholar
- Sarfstein R, Gorzalczany Y, Mizrahi A, Berdichevsky Y, Molshanski-Mor S, Weinbaum C, Hirshberg M, Dagher MC, Pick E: 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. Journal of Biological Chemistry. 2004, 279 (16): 16007-16016. 10.1074/jbc.M312394200.PubMedGoogle Scholar
- Han C-H, Freeman JLR, Lee T, Motalebi SA, Lambeth JD: Regulation of the neutrophil respiratory burst oxidase: Identification of an activation domain in p67phox. Journal of Biological Chemistry. 1998, 273: 16663-16668. 10.1074/jbc.273.27.16663.PubMedGoogle Scholar
- Nisimoto Y, Motalebi S, Han C-H, Lambeth JD: The p67phox activation domain regulates electron transfer flow from NADPH to flavin in flavocytochrome b558. Journal of Biological Chemistry. 1999, 274: 22999-23005. 10.1074/jbc.274.33.22999.PubMedGoogle Scholar
- Wilson MI, Gill DJ, Perisic O, Quinn MT, Williams RL: PB1 domain-mediated heterodimerization in NADPH oxidase and signaling complexes of atypical protein kinase C with Par6 and p62. Molecular Cell. 2003, 12 (1): 39-50. 10.1016/S1097-2765(03)00246-6.PubMedGoogle Scholar
- Kuribayashi F, Nunoi H, Wakamatsu K, Tsunawaki S, Sato K, Ito T, Sumimoto H: The adaptor protein p40(phox) as a positive regulator of the superoxide-producing phagocyte oxidase. The EMBO Journal. 2002, 21 (23): 6312-6320. 10.1093/emboj/cdf642.PubMed CentralPubMedGoogle Scholar
- Nakamura R, Sumimoto H, Mizuki K, Hata K, Ago T, Kitajima S, Takeshige K, Sakaki Y, Ito T: 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. 1998, 251 (3): 583-589. 10.1046/j.1432-1327.1998.2510583.x.PubMedGoogle Scholar
- Ellson CD, Davidson K, Ferguson GJ, O'Connor R, Stephens LR, Hawkins PT: Neutrophils from p40phox-/- mice exhibit severe defects in NADPH oxidase regulation and oxidant-dependent bacterial killing. Journal of Experimental Medicine. 2006, 203 (8): 1927-1937. 10.1084/jem.20052069.PubMed CentralPubMedGoogle Scholar
- Suh YA, Arnold RS, Lassegue B, Shi J, Xu X, Sorescu D, Chung AB, Griendling KK, Lambeth JD: Cell transformation by the superoxide-generating oxidase Mox1. Nature. 1999, 401 (6748): 79-82. 10.1038/43459.PubMedGoogle Scholar
- Bedard K, Krause KH: The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiological Reviews. 2007, 87 (1): 245-313. 10.1152/physrev.00044.2005.PubMedGoogle Scholar
- Banfi B, Clark RA, Steger K, Krause KH: Two novel proteins activate superoxide generation by the NADPH oxidase NOX1. Journal of Biological Chemistry. 2003, 278 (6): 3510-3513. 10.1074/jbc.C200613200.PubMedGoogle Scholar
- Geiszt M, Lekstrom K, Witta J, Leto TL: Proteins Homologous to p47phox and p67phox Support Superoxide Production by NAD(P)H Oxidase 1 in Colon Epithelial Cells. Journal of Biological Chemistry. 2003, 278 (22): 20006-20012. 10.1074/jbc.M301289200.PubMedGoogle Scholar
- Takeya R, Ueno N, Kami K, Taura M, Kohjima M, Izaki T, Nunoi H, Sumimoto H: Novel human homologues of p47phox and p67phox participate in activation of superoxide-producing NADPH oxidases. Journal of Biological Chemistry. 2003, 278 (27): 25234-25246. 10.1074/jbc.M212856200.PubMedGoogle Scholar
- Cheng G, Lambeth JD: NOXO1, Regulation of Lipid Binding, Localization, and Activation of Nox1 by the Phox Homology (PX) Domain. Journal of Biological Chemistry. 2004, 279 (6): 4737-4742. 10.1074/jbc.M305968200.PubMedGoogle Scholar
- Ueyama T, Geiszt M, Leto TL: Involvement of Rac1 in activation of multicomponent Nox1- and Nox3-based NADPH oxidases. Molecular and Cellular Biology. 2006, 26 (6): 2160-2174. 10.1128/MCB.26.6.2160-2174.2006.PubMed CentralPubMedGoogle Scholar
- Kawahara T, Kohjima M, Kuwano Y, Mino H, Teshima-Kondo S, Takeya R, Tsunawaki S, Wada A, Sumimoto H, Rokutan K: Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells. American Journal of Physiology – Cell Physiology. 2005, 288 (2): C450-457. 10.1152/ajpcell.00319.2004.PubMedGoogle Scholar
- Cheng G, Diebold BA, Hughes Y, Lambeth JD: Nox1-dependent reactive oxygen generation is regulated by Rac1. Journal of Biological Chemistry. 2006, 281 (26): 17718-17726. 10.1074/jbc.M512751200.PubMedGoogle Scholar
- Miyano K, Ueno N, Takeya R, Sumimoto H: Direct involvement of the small GTPase Rac in activation of the superoxide-producing NADPH oxidase Nox1. Journal of Biological Chemistry. 2006, 281 (31): 21857-21868. 10.1074/jbc.M513665200.PubMedGoogle Scholar
- Kawahara T, Kuwano Y, Teshima-Kondo S, Takeya R, Sumimoto H, Kishi K, Tsunawaki S, Hirayama T, Rokutan K: Role of nicotinamide adenine dinucleotide phosphate oxidase 1 in oxidative burst response to Toll-like receptor 5 signaling in large intestinal epithelial cells. The Journal of Immunology. 2004, 172 (5): 3051-3058.PubMedGoogle Scholar
- Yamamoto A, Kami K, Takeya R, Sumimoto H: Interaction between the SH3 domains and C-terminal proline-rich region in NADPH oxidase organizer 1 (Noxo1). Biochemical and Biophysical Research Communications. 2007, 352: 560-565. 10.1016/j.bbrc.2006.11.060.PubMedGoogle Scholar
- Banfi B, Malgrange B, Knisz J, Steger K, Dubois-Dauphin M, Krause KH: NOX3, a superoxide-generating NADPH oxidase of the inner ear. Journal of Biological Chemistry. 2004, 279 (44): 46065-46072. 10.1074/jbc.M403046200.PubMedGoogle Scholar
- Cheng G, Ritsick DR, Lambeth JD: Nox3 regulation by NOXO1, p47phox and p67phox. Journal of Biological Chemistry. 2004Google Scholar
- Kiss PJ, Knisz J, Zhang Y, Baltrusaitis J, Sigmund CD, Thalmann R, Smith RJ, Verpy E, Banfi B: Inactivation of NADPH oxidase organizer 1 results in severe imbalance. Current Biology. 2006, 16 (2): 208-213. 10.1016/j.cub.2005.12.025.PubMedGoogle Scholar
- Ueno N, Takeya R, Miyano K, Kikuchi H, Sumimoto H: The NADPH oxidase Nox3 constitutively produces superoxide in a p22phox-dependent manner: its regulation by oxidase organizers and activators. Journal of Biological Chemistry. 2005, 280 (24): 23328-23339. 10.1074/jbc.M414548200.PubMedGoogle Scholar
- Ambasta RK, Kumar P, Griendling KK, Schmidt HH, Busse R, Brandes RP: Direct interaction of the novel Nox proteins with p22phox is required for the formation of a functionally active NADPH oxidase. Journal of Biological Chemistry. 2004, 279 (44): 45935-45941. 10.1074/jbc.M406486200.PubMedGoogle Scholar
- Kawahara T, Ritsick D, Cheng G, Lambeth JD: Point mutations in the proline-rich region of p22phox are dominant inhibitors of Nox1- and Nox2-dependent reactive oxygen generation. Journal of Biological Chemistry. 2005, 280 (36): 31859-31869. 10.1074/jbc.M501882200.PubMedGoogle Scholar
- Martyn K, Frederick LM, von Loehneysen K, Dinauer MC, Knaus UG: Functional analysis of Nox4 reveals unique characteristics compared to other NADPH oxidases. Cellular Signalling. 2006, 18 (1): 69-82. 10.1016/j.cellsig.2005.03.023.PubMedGoogle Scholar
- Takemoto D, Tanaka A, Scott B: NADPH oxidases in fungi: Diverse roles of reactive oxygen species in fungal cellular differentiation. Fungal Genetics and Biology. 2007Google Scholar
- Lardy B, Bof M, Aubry L, Paclet M, Morel F, Satre M, Klein G: NADPH oxidase homologs are required for normal cell differentiation and morphogenesis in Dictyostelium discoideum. Biochimica et Biophysica Acta. 2005, 1744 (2): 199-212.PubMedGoogle Scholar
- Takemoto D, Tanaka A, Scott B: A p67Phox-like regulator is recruited to control hyphal branching in a fungal-grass mutualistic symbiosis. Plant Cell. 2006, 18 (10): 2807-2821. 10.1105/tpc.106.046169.PubMed CentralPubMedGoogle Scholar
- Aparicio S, Chapman J, Stupka E, Putnam N, Chia JM, Dehal P, Christoffels A, Rash S, Hoon S, Smit A: Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes. Science. 2002, 297 (5585): 1301-1310. 10.1126/science.1072104.PubMedGoogle Scholar
- Animal-Genome-Size-Database. [http://http//www.genomesize.com]
- Jaillon O, Aury JM, Brunet F, Petit JL, Stange-Thomann N, Mauceli E, Bouneau L, Fischer C, Ozouf-Costaz C, Bernot A: Genome duplication in the teleost fish Tetraodon nigroviridis reveals the early vertebrate proto-karyotype. Nature. 2004, 431 (7011): 946-957. 10.1038/nature03025.PubMedGoogle Scholar
- Inoue Y, Ogasawara M, Moroi T, Satake M, Azumi K, Moritomo T, Nakanishi T: Characteristics of NADPH oxidase genes (Nox2, p22, p47, and p67) and Nox4 gene expressed in blood cells of juvenile Ciona intestinalis. Immunogenetics. 2005, 57 (7): 520-534. 10.1007/s00251-005-0010-4.PubMedGoogle Scholar
- Sea Urchin Genome Sequencing Consortium, Sodergren EWG, Davidson EH, Cameron RA, Gibbs RA, Angerer RC, Angerer LM, Arnone MI, Burgess DR, Burke RD, Coffman JA, Dean M, Elphick MR, Ettensohn CA, Foltz KR, Hamdoun A, Hynes RO, Klein WH, Marzluff W, McClay DR, Morris RL, Mushegian A, Rast JP, Smith LC, Thorndyke MC, Vacquier VD, Wessel GM, Wray G, Zhang L, Elsik CG, Ermolaeva O, Hlavina W, Hofmann G, Kitts P, Landrum MJ, Mackey AJ, Maglott D, Panopoulou G, Poustka AJ, Pruitt K, Sapojnikov V, Song X, Souvorov A, Solovyev V, Wei Z, Whittaker CA, Worley K, Durbin KJ, Shen Y, Fedrigo O, Garfield D, Haygood R, Primus A, Satija R, Severson T, Gonzalez-Garay ML, Jackson AR, Milosavljevic A, Tong M, Killian CE, Livingston BT, Wilt FH, Adams N, Belle R, Carbonneau S, Cheung R, Cormier P, Cosson B, Croce J, Fernandez-Guerra A, Geneviere AM, Goel M, Kelkar H, Morales J, Mulner-Lorillon O, Robertson AJ, Goldstone JV, Cole B, Epel D, Gold B, Hahn ME, Howard-Ashby M, Scally M, Stegeman JJ, Allgood EL, Cool J, Judkins KM, McCafferty SS, Musante AM, Obar RA, Rawson AP, Rossetti BJ, Gibbons IR, Hoffman MP, Leone A, Istrail S, Materna SC, Samanta MP, Stolc V, Tongprasit W, Tu Q, Bergeron KF, Brandhorst BP, Whittle J, Berney K, Bottjer DJ, Calestani C, Peterson K, Chow E, Yuan QA, Elhaik E, Graur D, Reese JT, Bosdet I, Heesun S, Marra MA, Schein J, Anderson MK, Brockton V, Buckley KM, Cohen AH, Fugmann SD, Hibino T, Loza-Coll M, Majeske AJ, Messier C, Nair SV, Pancer Z, Terwilliger DP, Agca C, Arboleda E, Chen N, Churcher AM, Hallbook F, Humphrey GW, Idris MM, Kiyama T, Liang S, Mellott D, Mu X, Murray G, Olinski RP, Raible F, Rowe M, Taylor JS, Tessmar-Raible K, Wang D, Wilson KH, Yaguchi S, Gaasterland T, Galindo BE, Gunaratne HJ, Juliano C, Kinukawa M, Moy GW, Neill AT, Nomura M, Raisch M, Reade A, Roux MM, Song JL, Su YH, Townley IK, Voronina E, Wong JL, Amore G, Branno M, Brown ER, Cavalieri V, Duboc V, Duloquin L, Flytzanis C, Gache C, Lapraz F, Lepage T, Locascio A, Martinez P, Matassi G, Matranga V, Range R, Rizzo F, Rottinger E, Beane W, Bradham C, Byrum C, Glenn T, Hussain S, Manning G, Miranda E, Thomason R, Walton K, Wikramanayke A, Wu SY, Xu R, Brown CT, Chen L, Gray RF, Lee PY, Nam J, Oliveri P, Smith J, Muzny D, Bell S, Chacko J, Cree A, Curry S, Davis C, Dinh H, Dugan-Rocha S, Fowler J, Gill R, Hamilton C, Hernandez J, Hines S, Hume J, Jackson L, Jolivet A, Kovar C, Lee S, Lewis L, Miner G, Morgan M, Nazareth LV, Okwuonu G, Parker D, Pu LL, Thorn R, Wright R: The genome of the sea urchin Strongylocentrotus purpuratus. Science. 2006, 314 (5801): 941-952. 10.1126/science.1133609.PubMed CentralGoogle Scholar
- NCBI-Sea-Urchin-Genome-Database. [http://www.ncbi.nlm.nih.gov/genome/seq/BlastGen/BlastGen.cgi?taxid=7668]
- Hoegg S, Meyer A: Hox clusters as models for vertebrate genome evolution. Trends in Genetics. 2005, 21 (8): 421-424. 10.1016/j.tig.2005.06.004.PubMedGoogle Scholar
- Takeya R, Taura M, Yamasaki T, Naito S, Sumimoto H: Expression and function of Noxo1gamma, an alternative splicing form of the NADPH oxidase organizer 1. FEBS Journal. 2006, 273 (16): 3663-3677. 10.1111/j.1742-4658.2006.05371.x.PubMedGoogle Scholar
- Hata K, Ito T, Takeshige K, Sumimoto H: Anionic amphiphile-independent activation of the phagocyte NADPH oxidase in a cell-free system by p47 phox and p67 phox both in C terminally truncated forms. Journal of Biological Chemistry. 1998, 273: 4232-4236. 10.1074/jbc.273.7.4232.PubMedGoogle Scholar
- Nagasawa T, Ebisu K, Inoue Y, Miyano K, Tamura M: A new role of Pro-73 of p47phox in the activation of neutrophil NADPH oxidase. Archives of Biochemistry and Biophysics. 2003, 416 (1): 92-100. 10.1016/S0003-9861(03)00296-0.PubMedGoogle Scholar
- Ogura K, Nobuhisa I, Yuzawa S, Takeya R, Torikai S, Saikawa K, Sumimoto H, Inagaki F: NMR solution structure of the tandem Src homology 3 domains of p47phox complexed with a p22phox-derived proline-rich peptide. Journal of Biological Chemistry. 2006, 281 (6): 3660-3668. 10.1074/jbc.M505193200.PubMedGoogle Scholar
- Ago T, Kuribayashi F, Hiroaki H, Takeya R, Ito T, Kohda D, Sumimoto H: Phosphorylation of p47phox directs phox homology domain from SH3 domain toward phosphoinositides, leading to phagocyte NADPH oxidase activation. Proceedings of the National Academy of Sciences of the United States of America. 2003, 100 (8): 4474-4479. 10.1073/pnas.0735712100.PubMed CentralPubMedGoogle Scholar
- Finan P, Shimizu Y, Gout I, Hsuan J, Truong O, Butcher C, Bennett P, Waterfield MD, Kellie S: An SH3 domain and proline-rich sequence mediate an interaction between two components of the phagocyte NADPH oxidase complex. Journal of Biological Chemistry. 1994, 269 (19): 13752-13755.PubMedGoogle Scholar
- Leusen JH, Fluiter K, Hilarius PM, Roos D, Verhoeven AJ, Bolscher BG: 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. Journal of Biological Chemistry. 1995, 270 (19): 11216-11221. 10.1074/jbc.270.19.11216.PubMedGoogle Scholar
- Mizuki K, Takeya R, Kuribayashi F, Nobuhisa I, Kohda D, Nunoi H, Takeshige K, Sumimoto H: A region C-terminal to the proline-rich core of p47phox regulates activation of the phagocyte NADPH oxidase by interacting with the C-terminal SH3 domain of p67phox. Archives of Biochemistry and Biophysics. 2005, 444 (2): 185-194. 10.1016/j.abb.2005.10.012.PubMedGoogle Scholar
- Curwen V, Eyras E, Andrews TD, Clarke L, Mongin E, Searle SM, Clamp M: The Ensembl automatic gene annotation system. Genome Research. 2004, 14 (5): 942-950. 10.1101/gr.1858004.PubMed CentralPubMedGoogle Scholar
- Diebold BA, Bokoch GM: Molecular basis for Rac2 regulation of phagocyte NADPH oxidase. Nature Immunology. 2001, 2 (3): 211-215. 10.1038/85259.PubMedGoogle Scholar
- Leto T, Adams AG, de Mendez I: Assembly of the phagocyte NADPH oxidase: binding of Src homology 3 domains to proline-rich targets. The Proceedings of the National Academy of Sciences Online of the United State of America. 1994, 91 (22): 10650-10654. 10.1073/pnas.91.22.10650.Google Scholar
- Ito T, Matsui Y, Ago T, Ota K, Sumimoto H: Novel modular domain PB1 recognizes PC motif to mediate functional protein-protein interactions. The EMBO Journal. 2001, 20 (15): 3938-3946. 10.1093/emboj/20.15.3938.PubMed CentralPubMedGoogle Scholar
- Gauss KA, Bunger PL, Siemsen DW, Young CJ, Nelson-Overton L, Prigge JR, Swain SD, Quinn MT: Molecular analysis of the bison phagocyte NADPH oxidase: cloning and sequencing of five NADPH oxidase cDNAs. Comparative Biochemistry & Physiology Part B, Biochemistry & Molecular Biology. 2002, 133 (1): 1-12. 10.1016/S1096-4959(02)00090-8.Google Scholar
- Gauss KA, Mascolo PL, Siemsen DW, Nelson LK, Bunger PL, Pagano PJ, Quinn MT: Cloning and sequencing of rabbit leukocyte NADPH oxidase genes reveals a unique p67(phox) homolog. Journal of Leukocyte Biology. 2002, 71 (2): 319-328.PubMedGoogle Scholar
- Zhan Y, Virbasius JV, Song X, Pomerleau DP, Zhou GW: The p40phox and p47phox PX domains of NADPH oxidase target cell membranes via direct and indirect recruitment by phosphoinositides. Journal of Biological Chemistry. 2002, 277 (6): 4512-4518. 10.1074/jbc.M109520200.PubMedGoogle Scholar
- Stahelin RV, Burian A, Bruzik KS, Murray D, Cho W: Membrane binding mechanisms of the PX domains of NADPH oxidase p40phox and p47phox. Journal of Biological Chemistry. 2003, 278 (16): 14469-14479. 10.1074/jbc.M212579200.PubMedGoogle Scholar
- Ueyama T, Tatsuno T, Kawasaki T, Tsujibe S, Shirai Y, Sumimoto H, Leto T, Saito N: A Regulated Adaptor Function of p40phox: Distinct p67phox Membrane Targeting by p40phox and by p47phox. Molecular and Cellular Biology. 2007, 18 (2): 441-454.Google Scholar
- Honbou K, Minakami R, Yuzawa S, Takeya R, Suzuki NN, Kamakura S, Sumimoto H, Inagaki F: Full-length p40phox structure suggests a basis for regulation mechanism of its membrane binding. EMBO Journal. 2007, 26 (4): 1176-1186. 10.1038/sj.emboj.7601561.PubMed CentralPubMedGoogle Scholar
- Taylor R, Burritt J, Baniulis D, Foubert T, Lord C, Dinauer MC, Parkos CA, Jesaitis AJ: Site-specific inhibitors of NADPH oxidase activity and structural probes of flavocytochrome b: characterization of six monoclonal antibodies to the p22phox subunit. The Journal of Immunology. 2004, 173 (12): 7349-7357.PubMedGoogle Scholar
- Varnai P, Thyagarajan B, Rohacs T, Balla T: Rapidly inducible changes in phosphatidylinositol 4,5-bisphosphate levels influence multiple regulatory functions of the lipid in intact living cells. Journal of Cell Biology. 2006, 175 (3): 377-382. 10.1083/jcb.200607116.PubMed CentralPubMedGoogle Scholar
- Heo WD, Inoue T, Park WS, Kim ML, Park BO, Wandless TJ, Meyer T: PI(3,4,5)P3 and PI(4,5)P2 lipids target proteins with polybasic clusters to the plasma membrane. Science. 2006, 314 (5804): 1458-1461. 10.1126/science.1134389.PubMed CentralPubMedGoogle Scholar
- Ueyama T, Eto M, Kami K, Tatsuno T, Kobayashi T, Shirai Y, Lennartz MR, Takeya R, Sumimoto H, Saito N: Isoform-specific membrane targeting mechanism of Rac during Fc gamma R-mediated phagocytosis: positive charge-dependent and independent targeting mechanism of Rac to the phagosome. Journal of Immunology. 2005, 175 (4): 2381-2390.Google Scholar
- PROSITE-database. [http://ca.expasy.org/prosite/]
- Suh CI, Stull ND, Li XJ, Tian W, Price MO, Grinstein S, Yaffe MB, Atkinson S, Dinauer MC: The phosphoinositide-binding protein p40phox activates the NADPH oxidase during FcgammaIIA receptor-induced phagocytosis. Journal of Experimental Medicine. 2006, 203 (8): 1915-1925. 10.1084/jem.20052085.PubMed CentralPubMedGoogle Scholar
- Inoue Y, Suenaga Y, Yoshiura Y, Moritomo T, Ototake M, Nakanishi T: Molecular cloning and sequencing of Japanese pufferfish (Takifugu rubripes) NADPH oxidase cDNAs. Developmental & Comparative Immunology. 2004, 28 (9): 911-925. 10.1016/j.dci.2004.03.002.Google Scholar
- Halanych KM: The new view of animal phylogeny. Annual Review of Ecology, Evolution, and Systematics. 2004, 35: 229-256. 10.1146/annurev.ecolsys.35.112202.130124.Google Scholar
- Li S: Specificity and versatility of SH3 and other proline-recognition domains: structural basis and implication for cellular signal transduction. Biochemical Journal. 2005, 390: 641-653. 10.1042/BJ20050315.PubMed CentralPubMedGoogle Scholar
- Miller DJ, Hemmrich G, Ball EE, Hayward DC, Khalturin K, Funayama N, Agata K, Bosch TCG: The innate immune repertoire in Cnidaria-ancestral complexity and stochastic gene loss. Genome Biology. 2007, 8 (4): R59.51-R59.13.Google Scholar
- James-Clark H: Note on the infusoria flagellata and the spongiae ciliatae. American Journal of Science. 1866, 1: 113-114.Google Scholar
- Leadbeater BS, Kelly M: Evolution of animals-choanoflagellates and sponges. Water and Atmosphere. 2001, 9: 9-11.Google Scholar
- Snell EA, Furlong RF, Holland PWH: Hsp70 sequences indicate that choanoflagellates are closely related to animals. Current Biology. 2001, 11: 967-970. 10.1016/S0960-9822(01)00275-5.PubMedGoogle Scholar
- King N, Carroll SB: A receptor tyrosine kinase from choanoflagellates: molecular insights into early animal evolution. Proceedings of the National Academy of Sciences of the United States of America. 2001, 98 (26): 15032-15037. 10.1073/pnas.261477698.PubMed CentralPubMedGoogle Scholar
- King N, Hittinger CT, Carroll SB: Evolution of key cell signaling and adhesion protein family predates animal origins. Science. 2003, 301: 361-363. 10.1126/science.1083853.PubMedGoogle Scholar
- King N: The unicellular ancestry of animal development. Developmental Cell. 2004, 7: 313-325. 10.1016/j.devcel.2004.08.010.PubMedGoogle Scholar
- Lalucque H, Silar P: NADPH oxidase: an enzyme for multicellularity. TRENDS in Microbiology. 2003, 11 (1): 9-12. 10.1016/S0966-842X(02)00007-0.PubMedGoogle Scholar
- Cavalier-Smith T: Only six Kingdoms of life. Proceedings Royal Society of London B. 2004, 271: 1251-1262. 10.1098/rspb.2004.2705.Google Scholar
- Seals DF, Azucena EF, Pass I, Tesfay L, Gordon R, Woodrow M, Resau JH, Courtneidge SA: The adaptor protein Tks5/Fish is required for podosome formation and function, and for the protease-driven invasion of cancer cells. Cancer Cell. 2005, 7 (2): 155-165. 10.1016/j.ccr.2005.01.006.PubMedGoogle Scholar
- Malinin NL, Wright S, Seubert P, Schenk D, Griswold-Prenner I: Amyloid-beta neurotoxicity is mediated by FISH adapter protein and ADAM12 metalloprotease activity. Proceedings of the National Academy of Sciences of the United States of America. 2005, 102 (8): 3058-3063. 10.1073/pnas.0408237102.PubMed CentralPubMedGoogle Scholar
- NCBI-BLAST. [http://http//www.ncbi.nlm.nih.gov/BLAST/]
- Stahelin RV, Karathanassis D, Murray D, Williams RL, Cho W: Structural and Membrane Binding Analysis of the Phox Homology Domain of Bem1p: BASIS OF PHOSPHATIDYLINOSITOL 4-PHOSPHATE SPECIFICITY. Journal of Biological Chemistry. 2007, 282 (35): 25737-25747. 10.1074/jbc.M702861200.PubMedGoogle Scholar
- France YE, Boyd C, Coleman J, Novick PJ: The polarity-establishment component Bem1p interacts with the exocyst complex through the Sec15p subunit. Journal of Cell Science. 2006, 119 (Pt 5): 876-888. 10.1242/jcs.02849.PubMedGoogle Scholar
- Ignoul S, Eggermont J: CBS domains: structure, function, and pathology in human proteins. American Journal of Physiology – Cell Physiology. 2005, 289 (6): C1369-1378. 10.1152/ajpcell.00282.2005.PubMedGoogle Scholar
- Scott JW, Hawley SA, Green KA, Anis M, Stewart G, Scullion GA, Norman DG, Hardie DG: CBS domains form energy-sensing modules whose binding of adenosine ligands is disrupted by disease mutations. The Journal of Clinical Investigation. 2004Google Scholar
- Chung CY, Reddy TB, Zhou K, Firtel RA: A novel, putative MEK kinase controls developmental timing and spatial patterning in Dictyostelium and is regulated by ubiquitin-mediated protein degradation. Genes and Development. 1998, 12 (22): 3564-3578.PubMed CentralPubMedGoogle Scholar
- Kobayashi M, Ohura I, Kawakita K, Yokota N, Fujiwara M, Shimamoto K, Doke N, Yoshioka H: Calcium-dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase. Plant Cell. 2007, 19 (3): 1065-1080. 10.1105/tpc.106.048884.PubMed CentralPubMedGoogle Scholar
- Jagnandan D, Church JE, Banfi B, Stuehr DJ, Marrero MB, Fulton DJ: Novel mechanism of activation of NADPH oxidase 5. calcium sensitization via phosphorylation. Journal of Biological Chemistry. 2007, 282 (9): 6469-6507.Google Scholar
- Serrander L, Jaquet V, Bedard K, Plastre O, Hartley O, Arnaudeau S, Demaurex N, Schlegel W, Krause KH: NOX5 is expressed at the plasma membrane and generates superoxide in response to protein kinase C activation. Biochimie. 2007, 89 (9): 1159-1167. 10.1016/j.biochi.2007.05.004.PubMedGoogle Scholar
- Carol RJ, Dolan L: The role of reactive oxygen species in cell growth: lessons from root hairs. Journal of Experimental Botany. 2006, 57 (8): 1829-1834. 10.1093/jxb/erj201.PubMedGoogle Scholar
- Keller T, Damude HG, Werner D, Doerner P, Dixon RA, Lamb C: A plant homolog of the neutrophil NADPH oxidase gp91phox subunit gene encodes a plasma membrane protein with Ca2+ binding motifs. Plant Cell. 1998, 10 (2): 255-266. 10.1105/tpc.10.2.255.PubMed CentralPubMedGoogle Scholar
- Groom QJ, Torres MA, Fordham-Skelton AP, Hammond-Kosack KE, Robinson NJ, Jones JD: rbohA, a rice homologue of the mammalian gp91phox respiratory burst oxidase gene. The Plant Journal. 1996, 3: 515-22. 10.1046/j.1365-313X.1996.10030515.x.Google Scholar
- TIGR-Arabidopsis-thaliana-Protein-Database. [http://tigrblast.tigr.org/er-blast/index.cgi?project=ath1]
- Sumimoto H, Kamakura S, Ito T: Structure and function of the PB1 domain, a protein interaction module conserved in animals, fungi, amoebas, and plants. Sci STKE. 2007, 401:Google Scholar
- NCBI-HomoloGene. [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=homologene]
- Herve C, Tonon T, Collen J, Corre E, Boyen C: NADPH oxidases in Eukaryotes: red algae provide new hints!. Current Genetics. 2006, 49 (3): 190-204. 10.1007/s00294-005-0044-z.PubMedGoogle Scholar
- NCBI-Fungi-Genomes-BLAST-server. [http://www.ncbi.nlm.nih.gov/sutils/genom_table.cgi?organism=fungi]
- JGI-Genomes-Database. [http://genome.jgi-psf.org/euk_home.html]
- Broad-Institute-Genomes-database. [http://www.broad.mit.edu/cgi-bin/annotation/fgi/blast_page.cgi]
- Pfam-search. [http://www.sanger.ac.uk/Software/Pfam/]
- ClustalW. [http://http//www.ddbj.nig.ac.jp/search/clustalw-j.html]
- Ensembl-AlignSliceView-program. [http://www.ensembl.org/Homo_sapiens/alignsliceview]
- Hedges SB: The origin and evolution of model organisms. Nature Reviews Genetics. 2002, 3 (11): 838-849. 10.1038/nrg929.PubMedGoogle Scholar
- Sumimoto H, Hata K, Mizuki K, Ito T, Kage Y, Sakaki Y, Fukumaki Y, Nakamura M, Takeshige K: Assembly and activation of the phagocyte NADPH oxidase: Specific Interaction of the N-terminal Src homology 3 domain of p47phox with p22phox is required for activation of the NADPH oxidase complex. Journal of Biological Chemistry. 1996, 36: 22152-22158.Google Scholar
- Sumimoto H, Kage Y, Nunoi H, Sasaki H, Nose T, Fukumaki Y, Ohno M, Minakami S, Takeshige K: Role of Src homology 3 domains in assembly and activation of the phagocyte NADPH oxidase. Proceedings of the National Academy of Sciences of the United States of America. 1994, 91: 5345-5349. 10.1073/pnas.91.12.5345.PubMed CentralPubMedGoogle Scholar
- Nobuhisa I, Takeya R, Ogura K, Ueno N, Kohda D, Inagaki F, Sumimoto H: Activation of the superoxide-producing phagocyte NADPH oxidase requires co-operation between the tandem SH3 domains of p47phox in recognition of a polyproline type II helix and an adjacent alpha-helix of p22phox. Biochemical Journal. 2006, 396 (1): 183-192. 10.1042/BJ20051899.PubMed CentralPubMedGoogle Scholar
- Edens WA, Sharling L, Cheng G, Shapira R, Kinkade JM, Edens HA, Tang X, Flaherty DB, Benian G, Lambeth JD: Tyrosine cross-linking of extracellullar matrix is catalyzed by Duox, a multidomain oxidase/peroxidase with homology to the phagocyte oxidase subunit gp91phox. Journal of Cell Biology. 2001, 154: 879-891. 10.1083/jcb.200103132.PubMed CentralPubMedGoogle Scholar
- Ha EM, Oh CT, Bae YS, Lee WJ: A direct role for dual oxidase in Drosophila gut immunity. Science. 2005, 310 (5749): 847-850. 10.1126/science.1117311.PubMedGoogle Scholar
- Ritsick DR, Edens WA, Finnerty V, Lambeth JD: Nox regulation of smooth muscle contraction. Free Radical Biology & Medicine. 2007, 43 (1): 31-38. 10.1016/j.freeradbiomed.2007.03.006.Google Scholar
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