• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1455-1463
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• 2014

In the past, reactive oxygen species (ROS) have been considered a harmful byproduct of aerobic metabolism. However, accumulating evidence implicates redox homeostasis, which maintains appropriate ROS levels, in cell proliferation and differentiation in plants and animals. Similarly, ROS generation and signaling are instrumental in fungal development and host-fungus interaction. In fungi, NADPH oxidase, a homolog of human $gp91^{phox}$, generates superoxide and is the main source of ROS. The mechanism of activation and signaling by NADPH oxidases in fungi appears to be largely comparable to those in plants and animals. Recent studies have shown that the fungal NADPH oxidase homologs NoxA (Nox1), NoxB (Nox2), and NoxC (Nox3) have distinct functions. In particular, these studies have consistently demonstrated the impact of NoxA on the development of fungal multicellular structures. Both NoxA and NoxB (but not NoxC) are involved in host-fungus interactions, with the function of NoxA being more critical than that of NoxB.

• Biomolecules & Therapeutics
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• v.28 no.1
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• pp.25-33
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• 2020

Several recent studies have reported that reactive oxygen species (ROS), superoxide anion and hydrogen peroxide (H₂O₂), play important roles in various cellular signaling networks. NADPH oxidase (Nox) isozymes have been shown to mediate receptor-mediated ROS generation for physiological signaling processes involved in cell growth, differentiation, apoptosis, and fibrosis. Detectable intracellular levels of ROS can be induced by the electron leakage from mitochondrial respiratory chain as well as by activation of cytochrome p450, glucose oxidase and xanthine oxidase, leading to oxidative stress. The up-regulation and the hyper-activation of NADPH oxidases (Nox) also likely contribute to oxidative stress in pathophysiologic stages. Elevation of the renal ROS level through hyperglycemia-mediated Nox activation results in the oxidative stress which induces a damage to kidney tissues, causing to diabetic nephropathy (DN). Nox inhibitors are currently being developed as the therapeutics of DN. In this review, we summarize Nox-mediated ROS generation and development of Nox inhibitors for therapeutics of DN treatment.

• Natural Product Sciences
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• v.24 no.1
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• pp.59-65
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• 2018

An isoform of NADPH oxidase (NOX), NOX2 is a superoxide-generating enzyme involved in diverse pathophysiological events. Although its potential as a therapeutic target has been validated, there is no clinically available inhibitor. Herein, NOX2-inhibitory activity was screened with the constituents isolated from Schisandra chinensis, which has been reported to have antioxidant and reactive oxygen species (ROS)-scavenging effects. Among the partitions prepared from crude methanolic extract, a chloroform-soluble partition showed the highest NOX2-inhibitory activity in PLB-985 cell-based NOX2 assay. A total of twenty nine compounds (1 - 29) were identified from the chloroform fraction, including two first isolated compounds; dimethyl-malate (25) and 2-(2-hydroxyacetyl) furan (27) from this plants. Of these constituents, two compounds (gomisin T, and pregomisin) exhibited an NOX2-inhibitory effect with the $IC_{50}$ of $9.4{\pm}3.6$, and $62.9{\pm}11.3{\mu}M$, respectively. They are confirmed not to be nonspecific superoxide scavengers in a counter assay using a xanthine-xanthine oxidase system. These findings suggest the potential application of gomisin T (6) and other constituents of S. chinensis to inhibit NOX2.

• Mycobiology
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• v.42 no.3
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• pp.241-248
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• 2014

NADPH oxidases (Noxes), transmembrane proteins found in most eukaryotic species, generate reactive oxygen species and are thereby involved in essential biological processes. However, the fact that genes encoding ferric reductases and ferric-chelate reductases share high sequence similarities and domains with Nox genes represents a challenge for bioinformatic approaches used to identify Nox-encoding genes. Further, most studies on fungal Nox genes have focused mainly on functionality, rather than sequence properties, and consequently clear differentiation among the various Nox isoforms has not been achieved. We conducted an extensive sequence analysis to identify putative Nox genes among 34 eukaryotes, including 28 fungal genomes and one Oomycota genome. Analyses were performed with respect to phylogeny, transmembrane helices, di-histidine distance and glycosylation. Our analyses indicate that the sequence properties of fungal Nox genes are different from those of human and plant Nox genes, thus providing novel insight that will enable more accurate identification and characterization of fungal Nox genes.

• Toxicological Research
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• v.30 no.3
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• pp.149-157
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• 2014

Smoking is one of the most serious but preventable causes of cardiovascular disease (CVD). Key aspects of pathological process associated with smoking include endothelial dysfunction, a prothrombotic state, inflammation, altered lipid metabolism, and hypoxia. Multiple molecular events are involved in smoking-induced CVD. However, the dysregulations of reactive oxygen species (ROS) generation and metabolism mainly contribute to the development of diverse CVDs, and NADPH oxidase (NOX) has been established as a source of ROS responsible for the pathogenesis of CVD. NOX activation and resultant ROS production by cigarette smoke (CS) treatment have been widely observed in isolated blood vessels and cultured vascular cells, including endothelial and smooth muscle cells. NOX-mediated oxidative stress has also been demonstrated in animal studies. Of the various NOX isoforms, NOX2 has been reported to mediate ROS generation by CS, but other isoforms were not tested thoroughly. Of the many CS constituents, nicotine, methyl vinyl ketone, and ${\alpha}$,${\beta}$-unsaturated aldehydes, such as, acrolein and crotonaldehyde, appear to be primarily responsible for NOX-mediated cytotoxicity, but additional validation will be needed. Human epidemiological studies have reported relationships between polymorphisms in the CYBA gene encoding p22phox, a catalytic subunit of NOX and susceptibility to smoking-related CVDs. In particular, G allele carriers of A640G and $-930^{A/G}$ polymorphisms were found to be vulnerable to smoking-induced cardiovascular toxicity, but results for C242T studies are conflicting. On the whole, evidence implicates the etiological role of NOX in smoking-induced CVD, but the clinical relevance of NOX activation by smoking and its contribution to CVD require further validation in human studies. A detailed understanding of the role of NOX would be helpful to assess the risk of smoking to human health, to define high-risk subgroups, and to develop strategies to prevent or treat smoking-induced CVD.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2984-2988
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• 2009

A gene (PH0311) encoding a hypothetical protein from the genome sequence data of the hyperthermophilic archaeon Pyrococcus horikoshii OT3 was cloned and over-expressed in Escherichia coli. The purified recombinant protein was found to possess FAD-dependent NADH oxidase activity, although it lacked sequence homology to any other known general NADH oxidase family. The product of the PH0311 gene was thus designated PhNOX (NADH oxidase from Pyrococcus horikoshii), with an estimated molecular weight of 84 kDa by gel filtration and 22 kDa by SDS-PAGE, indicating it to be a homotetramer of 22 kDa subunits. PhNOX catalyzed the oxidation of reduced ${\beta}$-NADH with subsequent formation of $H_2O_2$ in the presence of FAD as a cofactor, but not ${\alpha}$-NADH, ${\alpha}$-NADPH, or ${\beta}$-NADPH. PhNOX showed high affinity for ${\beta}$-NADH with a Km value of 3.70 ${\mu}$M and exhibited optimum activity at pH 8.0 and 95$^{\circ}C$ as it is highly stable against high temperature.

• The Plant Pathology Journal
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• v.38 no.4
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• pp.345-354
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• 2022

NADPH oxidase (Nox) complexes are known to play essential roles in differentiation and proliferation of many filamentous fungi. However, the functions of Noxs have not been elucidated in Colletotrichum species. Therefore, we set out to characterize the roles of Nox enzymes and their regulators in Colletotrichum scovillei, which causes serious anthracnose disease on pepper fruits in temperate and subtropical and temperate region. In this study, we generated targeted deletion mutants for CsNox1, CsNox2, CsNoxR, and CsNoxD via homologous recombination. All deletion mutants were normal in mycelial growth, conidiation, conidial germination, and appressorium formation, suggesting that CsNox1, CsNox2, CsNoxR, and CsNoxD are not involved in those developmental processes. Notably, conidia of 𝜟Csnox2 and 𝜟Csnoxr, other than 𝜟Csnox1 and 𝜟Csnoxd, failed to cause anthracnose on intact pepper fruits. However, they still caused normal disease on wounded pepper fruits, suggesting that Csnox2 and CsnoxR are essential for penetration-related morphogenesis in C. scovillei. Further observation proved that 𝜟Csnox2 and 𝜟Csnoxr were unable to form penetration peg, while they fully developed appressoria, revealing that defect of anthracnose development by 𝜟Csnox2 and 𝜟Csnoxr resulted from failure in penetration peg formation. Our results suggest that CsNox2 and CsNoxR are critical for appressorium-mediated penetration in C. scovillei-pepper fruit pathosystem, which provides insight into understanding roles of Nox genes in anthracnose disease development.

• IMMUNE NETWORK
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• v.23 no.5
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• pp.42.1-42.21
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• 2023

When the lungs are infected with bacteria, alveolar macrophages (AMs) are recruited to the site and play a crucial role in protecting the host by reducing excessive lung inflammation. However, the regulatory mechanisms that trigger the recruitment of AMs to lung alveoli during an infection are still not fully understood. In this study, we identified a critical role for NADPH oxidase 4 (NOX4) in the recruitment of AMs during Staphylococcus aureus lung infection. We found that NOX4 knockout (KO) mice showed decreased recruitment of AMs and increased lung neutrophils and injury in response to S. aureus infection compared to wildtype (WT) mice. Interestingly, the burden of S. aureus in the lungs was not different between NOX4 KO and WT mice. Furthermore, we observed that depletion of AMs in WT mice during S. aureus infection increased the number of neutrophils and lung injury to a similar level as that observed in NOX4 KO mice. Additionally, we found that expression of intercellular adhesion molecule-1 (ICAM1) in NOX4 KO mice-derived lung endothelial cells was lower than that in WT mice-derived endothelial cells. Therefore, we conclude that NOX4 plays a crucial role in inducing the recruitment of AMs by controlling ICAM1 expression in lung endothelial cells, which is responsible for resolving lung inflammation during acute S. aureus infection.

• Journal of Periodontal and Implant Science
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• v.36 no.4
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• pp.863-878
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• 2006

Porphyromonas gingivalis는 치주질환을 야기하는 독성세균으로서, 구강상피세포에 p. gingivalis가 감염되었을 때, 세포형태에 변화를 초래함으로 인해 방어기작이 작동하게 된다. 치주질환과 관련되어 생성된 활성 산소종의 소거에 관여하는 항산화성분은 p. gingivalis 이 감염된 구강상피세포에서 그 분포와 발현수준이 달라지리라 예상된다. 따라서 이번 연구에서는 구강상피세포(KB 세포)에 p. gingivalis가 감염되었을 때 야기되는 활성산소종과 이를 소거하는 역할을 하는 항산화단백들의 역할들을 규명하고자 하였다. 활성산소종 형성을 조절하는 NADPH oxidase 중 NOX4와 Rac1 전사체는 구강상피세포에서 p. gingivalis세균에 의해 증가하였으며 $gp91^{phox}$, Rac2, $p47^{phox}$와 $p67^{phox}$는 세균에 의한 변화가 관찰되지 않았다. 반면에 $p40^{phox}$ 전사체는 감소하는 경향을 보였다. NOX1 전사체는 p. gingivalis 처리 30분 후 감소하였다가 60분 후에는 다시 증가하는 양상을 보였다. 같은 시간에 NOX 활성화 단백인 NOXA1은 감소하고, NOX 구성단백질인 NOXO1은 증가하는 경향을 보였다. p. gingivalis가 감염된 구강상피세포를 방어하는 항산화단백 발현수준을 평가한 결과, SOD1, 2, 3 모두 p. gingivalis 처리시간에 따라 증가하는 양상을 보였다. GPx 발현 양상도 SOD와 유사하게 나타났다. $H_2O_2$를 소거하는 Prx는 감염된 KB 세포에서 Prx4와 Prx5가 4-6배 증가하는 것을 알 수 있었다. 반면 endocytosis 과정 중 $H_2O_2$ 생산은 변화되지 않았다. 이번 연구의 결과, p. gingivalis의 감염은 KB 세포의 NOX4와 Rac1의 NADPH oxidase 발현을 증가시켰으며, NOX1은 NOXA1과 NOXO1의 조절에 의해 영향을 받음을 알 수 있었다. 또한 항산화기작으로는 SOD, GPx, Prx가 증가하였는데, 이것은 Prx4와 Prx5가 중요한 역할을 할 것을 시사하였다.

• Parasites, Hosts and Diseases
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• v.51 no.1
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• pp.61-68
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• 2013

Entamoeba histolytica, which causes amoebic colitis and occasionally liver abscess in humans, is able to induce host cell death. However, signaling mechanisms of colon cell death induced by E. histolytica are not fully elucidated. In this study, we investigated the signaling role of NOX in cell death of HT29 colonic epithelial cells induced by E. histolytica. Incubation of HT29 cells with amoebic trophozoites resulted in DNA fragmentation that is a hallmark of apoptotic cell death. In addition, E. histolytica generate intracellular reactive oxygen species (ROS) in a contact-dependent manner. Inhibition of intracellular ROS level with treatment with DPI, an inhibitor of NADPH oxidases (NOXs), decreased Entamoebainduced ROS generation and cell death in HT29 cells. However, pan-caspase inhibitor did not affect E. histolytica-induced HT29 cell death. In HT29 cells, catalytic subunit NOX1 and regulatory subunit Rac1 for NOX1 activation were highly expressed. We next investigated whether NADPH oxidase 1 (NOX1)-derived ROS is closely associated with HT29 cell death induced by E. histolytica. Suppression of Rac1 by siRNA significantly inhibited Entamoeba-induced cell death. Moreover, knockdown of NOX1 by siRNA, effectively inhibited E. histolytica-triggered DNA fragmentation in HT29 cells. These results suggest that NOX1-derived ROS is required for apoptotic cell death in HT29 colon epithelial cells induced by E. histolytica.