• Title/Summary/Keyword: NO synthase

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Expression in Escherichia coli of a Putative Human Acetohydroxyacid Synthase

  • Duggleby, Ronald G.;Kartikasari, Apriliana E.R.;Wunsch, Rebecca M.;Lee, Yu-Ting;Kil, Mee-Wha;Shin, Ju-Young;Chang, Soo-Ik
    • BMB Reports
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    • v.33 no.3
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    • pp.195-201
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    • 2000
  • A human gene has been reported that may encode the enzyme acetohydroxyacid synthase. Previously this enzyme was thought to be absent from animals although it is present in plants and many microorganisms. In plants, this enzyme is the target of a number of commercial herbicides and the use of these compounds may need to be reassessed if the human enzyme exists and proves to be susceptible to inhibition. Here we report the construction of several plasmid vectors containing the cDNA sequence for this protein, and their expression in Escherichia coli. High levels of expression were observed, but most of the protein proved to be insoluble. The small amounts of soluble protein contained little or no acetohydroxyacid synthase activity. Attempts to refold the insoluble protein were successful insofar as the protein became soluble. However, the refolded protein did not gain any acetohydroxyacid synthase activity. In vivo complementation tests of an E. coli mutant produced no evidence that the protein is active. Incorrect folding, or the lack of another subunit, may explain the data but we favor the interpretation that this gene does not encode an acetohydroxyacid synthase.

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Inhibition of Nitric Oxide Synthesis by 8-epi-xanthatin in Activated RAW 264.7 Cells (활성화한 RAW 264.7 세포주에서 8-epi-xanthatin의 Nitric Oxide 생성저해)

  • Lee, Hwa-Jin;Jeong, Yeon-Su;Ryu, Shi-Yong;Ryu, Jae-Ha
    • YAKHAK HOEJI
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    • v.42 no.5
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    • pp.540-543
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    • 1998
  • The nitric oxide (NO) produced in large amounts by inducible nitric oxide synthase is known to be responsible for the vasodilation and hypotension observed in septic shock. We have found that 8-epi-xanthatin from Xanthium strumarium L. inhibited the production of NO in LPS-activated RAW 264.7 cells ($IC_{50}$ value was 1.5 ${\mu}$M). This activity was resulted from the suppressing of inducible nitric oxide synthase enzyme expression.

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Elicitor-treated extracts of Saururus chinensis inhibit the expression of inducible nitric oxide synthase and cyclooxygenase-2 enzyme expression in Raw cells for suppression of inflammation

  • Lee, Eun-Ho;Park, Hye-Jin;Kim, Dong-Hee;Jung, Hee-Young;Kang, In-Kyu;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.149-155
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    • 2019
  • Elicitor treatment was performed to increase the anti-inflammatory activity of useful biological sources. The result showed that elicitor-treated Saururus chinensis leaf extracts positively affected nitric oxide (NO) production, and the expression of inducible NO synthase and cyclooxygenase-2 compared to extracts not exposed to elicitor treatment. This finding identified elicitor treatment as a suitable strategy for increasing the biological activity of S. chinensis. Therefore, elicitor-treated S. chinensis is useful both as health functional and medicinal materials.

Glyphosate Toxicity: II. EPSP-synthase Activity in Cell Suspension Culture of Corydalis Sempervirens and Lycopersicon Esculentum (Glyphosate 독성(毒性): II. corydalis Sempervirens와 토마토의 세포배양체(細胞培養體)에서 EPSP-synthase의 활성(活性)에 미치는 영향(影響))

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.148-153
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    • 1995
  • Glyphosate (N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves or sprayed to the whole plants of tomato(Lycopersicon esculentum Mil var. Moneymaker) induced the rapid inhibition of 5-enolpyruvyl skimic acid 3-phosphate synthase(EPSP-synthase). It shows that EPSP-synthase activity precedes chlorophyll loss. There is no difference in EPSP-synthase activity between in vivo tomato meristem and cell suspension culture if glyphosate is not applied. The EPSP-synthase activity is in a range of 4 to 6 nkat per mg protein. The inhibition of EPSP-synthase action is induced within 36 h after glyphosate application while the Chl contents were reduced 48 h after the application. In cell suspension culture of tomato and Corydalis (Corydalis sempervirens), a sublethal concentration of glyphosate retards the fresh weight increase and prolonged lag phase. The fresh weight is reached maximal about 14 days after the subculture in the presence of glyphosate. The inhibitory effect of glyphosate on EPSP-synthase is remarkably induced in lag phase.

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Phenethyl Isothiocyanate Inhibits Ovalbumin-induced Inducible Nitric Oxide Synthase Expression (Ovalbumin에 의해서 유도된 inducible nitric oxide synthase 발현에 대한 phenethyl isothiocyanate의 억제효과)

  • Shin, Hwa-Jeong;Youn, Hyung-Sun
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.759-762
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    • 2012
  • Egg allergies have been reported as one of the most prevalent food hypersensitivities in the pediatric population. One of the major egg allergens is ovalbumin (OVA), which is the major protein in the egg whites. Phenethyl isothiocyanate (PEIC) from cruciferous vegetables has an effect on anti-inflammatory therapy. In the present report, we show that PEIC inhibits the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activation induced by OVA. PEIC also inhibits the OVA-induced inducible nitric oxide synthase (iNOS) expression and nitrite production. However, PEIC did not suppress the cyclooxygenase-2 (COX-2) expression induced by OVA. These results suggest that PEIC has the specific mechanism for anti-inflammatory responses and efficient anti-allergic activities.

Therapeutic Application of Nitric Oxide in Human Diseases

  • NamKoong, Seung;Kim, Young-Myeong
    • Biomolecules & Therapeutics
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    • v.18 no.4
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    • pp.351-362
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    • 2010
  • Nitric oxide (NO), synthesized from L-arginine by three isoforms of NO synthase (NOS), is a gaseous signaling molecule with an astonishingly wide range of biological and pathophysiological activities, including vasorelaxation, angiogenesis, anti-inflammation, and anti-apoptosis in mammalian cells. Recent studies have shown that NO donors and inhaled NO convert to biologically active NO under biological conditions and act as a signaling molecule in pathophysiological conditions. This review will discuss the roles of NO and its potential therapeutic implication in various human diseases, such as tumor, vascular regeneration, hypertension, wound healing, and ischemia-reperfusion injury.

Regulation of NO from Endothelial Cells by the Decrease of Cellular cAMP Under Arsenite Exposure

  • Lee, Soo-Youn;Min, Ji-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.392-395
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    • 2008
  • In an attempt to delineate the direct effect of arsenite-induced endothelial dysfunction on nitric oxide (NO) production, confluent bovine aortic endothelial cells (BAEC) were incubated with arsenite, and endothelial NO synthase expression and NO production were measured. Exposure of arsenite decreased NO production for up to 24h. This decrease was accompanied by decreases in cAMP, protein kinase A (PKA) activity, and furthermore, significant reduction of pCREB. In conclusion, this study is the first to demonstrate that exposure of arsenite decreases NO production by a reduction of pCREB and PKA activity that may be mediated by cAMP, leading to endothelial dysfunction.

Puerariae flos inhibits inflammatory responses in interferon-γ and lipopolysaccharide-stimulated mouse peritoneal macrophages

  • Hong, Seung-Heon;Kim, Hong-Joon;Cha, Dong-Seok;Lee, Ju-Young;Na, Ho-Jeong
    • Advances in Traditional Medicine
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    • v.7 no.3
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    • pp.229-234
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    • 2007
  • In macrophages, nitric oxide (NO) is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions including inflammation. In this study, we have examined the inhibition effects of NO production by 85% methanol extract of the flower of Pueraria thunbergiana (PF) in mouse macrophages. Extract of PF (1, 10, 100 ${\mu}g/ml$) inhibited NO production, inducible NO synthase and cyclooxygenase-2 expression in interferon-g and lipopolysaccharide-stimulated mouse peritoneal macrophages and it had no cytotoxicity. These data suggest that 85% methanol extract of PF might be useful in controlling macrophages mediated inflammatory disease.

The Effects of Diesel Exhaust Particles on the Alveolar Macrophages for Inducible Nitric Oxide Synthase Induction and Nitric Oxide with Nitrotyrosilated-protein Formation (디젤분진이 폐포대식세포에서 nitric oxide의 생성과 inducible nitric oxide synthase의 발현 및 nitrotyrosilated-protein의 형성에 미치는 효과)

  • Lim Young;Choe Myung-Ok;Lee Kweon-Haeng;Kim Kyung-A;Kim Kil-Soo;Lee Myoung-Heon;Li Tian-Zhu;Lee Soo-Jin;Choe Nong-Hoon
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.192-198
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    • 2006
  • Epidemiological studies have demonstrated an association between exposure to diesel exhaust particles (DEP) and adverse cardiopulmonary effects. Despite the epidemiological proof, the pathogenesis of DEP-related pulmonary diseases remain poorly understood. So, comprehensive in vivo and in vitro researches are required to know the effects of DEP on diverse lung diseases. Alveolar macrophages (AM) and airway epithelial cells are known as important cellular targets in DEP-induced lung diseases. Other studies have shown that nitric oxide (NO) is involved in particle matter induced lung injury. The present study was undertaken to determine whether DEP has an synergistic effects on lipopolysaccharide (LPS)-induced NO formation and inducible nitric oxide synthase (iNOS) with nitrotyrosilated-protein formation in cultured primary alveolar macrophages. The formation of NO was determined through the Griess reaction in the cultured medium and iNOS with nitrotyrosilated-proteins are analyzed by immunohistochemical staining and Western analysis. The results indicate that DEP exposure does not induce NO formation by itself, however DEP showed significant synergistic effects on LPS-induced NO formation. So, our results suggest that DEP inhalation could aggravate inflammatory lung disease through NO formation.

Inhibitory Effect of Galangin from Alpinia officinarum on Lipopolysaccharide-induced Nitric Oxide Synthesis in RAW 264.7 macrophages (고량강으로부터 분리된 galangin의 RAW 264.7 세포주에서 LPS로 유도된 nitric oxide 생성 저해활성)

  • Lee, Hwa Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.511-515
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    • 2014
  • In a screen for plant-derived inhibitors of nitric oxide (NO) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells, a flavonol isolated from the chloroform extract of Alpinia officinarum was isolated. The structure of the flavonol was found to be 3,5,7-trihydroxy-2-phenylchromen-4-one (galangin, GLG) by using spectroscopy. GLG exhibited an inhibitory effect ($IC_{50}$ value: $26.8{\mu}M$) on NO production in LPS-stimulated RAW 264.7 murine macrophage cells. Moreover, GLG suppressed expressions of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner.