Superoxide Generation by Nox1 in Guinea Pig Gastric Mucosal Cells Involves a Component with p67^<phox>-Ability(Biochemistry/Molecular Biology)
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概要
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Nox1, a homologue of gp91^<phox> subunit of the phagocyte NADPH oxidase, is responsible for spontaneous superoxide (O^-_2) generation in guinea pig gastric mucosal cells (GMC), but involvement of regulatory components (p67^<phox>, p47^<phox>, and Rac) which are essential in phagocytes remains unknown. Here, we aimed to figure out how Nox1 of GMC achieves an active oxidase status. CMC in primary culture show low O^-_2 generation but acquire a 9-fold higher activity when cultured with Helicobacter pylori lipopolysaccharide (LPS), in correlation with a far increased Nox1 expression. Investigation into the O^-_2-generating ability of LPS-induced Nox1 in cell-free reconstitution assays showed that: 1) Nox1 is unable to generate O^-\2 per se-; 2) the combination of Nox1 with GMC cytosol is insufficient for a significant O^-_2 generation; 3) the combination with neutrophil cytosol enables Nox1 to act like gp91^<phox>, i.e., to produce O^-_2 appreciably in response to myristate stimulation; 4) Nox1 prefers NADPH to NADH under the in vitro assay with neutrophil cytosol plus myristate (K_m=10.4 μM); 5) substitution of neutrophil cytosol by a set of recombinant cytosolic components (rp07^<phox>, rp47^<phox>, Rac2) is, however, ineffective and still requires GMC cytosol. Thus, Nox1 probably requires an additional cytosolic factor(s). In contrast, GMC cytosol enables cytochrome b_<558> to generate plenty of O^-_2, on condition that rp47^<phox> is added. This result suggests that GMC cytosol contains a component with p67^<phox>-ability, and also Rac, but lacks p47^<phox>. These data indicate that GMC Nox1 requires at least a p67^<phox> counterpart and Rac to acquire NADPH oxidase activity.
- 公益社団法人日本薬学会の論文
- 2004-02-01
著者
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ROKUTAN Kazuhito
Department of Nutrition, School of Medicine, University of Tokushima
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YOSHIDA Lucia
Department of Food Chemistry, Faculty of Agriculture, Tohoku University
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Rokutan K
Department Of Nutrition School Of Medicine University Of Tokushima
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Rokutan Kazuhito
Department Of Nutrition School Of Medicine University Of Tokushima
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Rokutan Kazuhito
Department Of Nutrition School Of Medicine The University Of Tokushima
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Yoshida Lucia
Department Of Food Chemistry Faculty Of Agriculture Tohoku University
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Yoshida Lucia
Department Of Infectious Diseases National Research Institute For Child Health And Development;
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SHIMOYAMA TAKASHI
Department of Gastroenterology, Hyogo College of Medicine
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KOBAYASHI Toshihiro
Department of Anatomy and Cell Biology
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Tsunawaki S
Department Of Infectious Diseases National Research Institute For Child Health And Development
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Kawahara T
Department Of Nutrition School Of Medicine University Of Tokushima
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NISHIDA Satoshi
Department of Infectious Diseases, National Research Institute for Child Health and Development;
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KAWAHARA Tsukasa
Department of Nutrition, School of Medicine, University of Tokushima
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KONDO-TESHIMA Shigetada
Department of Nutrition, School of Medicine, University of Tokushima
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TSUNAWAKI Shohko
Department of Infectious Diseases, National Research Institute for Child Health and Development;
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Kondo-teshima Shigetada
Department Of Nutrition School Of Medicine University Of Tokushima
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Kobayashi T
Department Of Anatomy And Cell Biology Kochi Medical School
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Kawahara T
Department Of Materials Science And Engineering National Defense Academy
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Kawahara Tsukasa
Department Of Nutrition School Of Medicine The University Of Tokushima
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Nishida S
Graduate School Of Pharmaceutical Sciences The Univ. Of Tokyo
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Tsunawaki Shohko
Department Of Infectious Diseases National Research Institute For Child Health And Development;
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Tsunawaki Shohko
Department Of Infections Disease National Children's Medical Research Center
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Kondo-Teshima S
Department of Nutrition, School of Medicine, University of Tokushima
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