• Title/Summary/Keyword: Heme protein

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Isolation and characterization of a noval membrane-bound cytochrome $C_{553}$ from the strictly anaerobic phototroph, heliobacillus mobilis

  • Lee, Woo-Yiel;Bla;Kim, Seung-Ho
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.206-212
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    • 1997
  • Heliobacillus mobilis is a strictly anaerobic Gram-positive bacterium which contains a primitive Photosystem I-type reaction center. The membrane-bound cytochrome $C_{553}$ from the heliobacterium suggested to be the immediate electron donor to the photooxidized pigment (P798+) has been isolated and characterized. The heme protein was visualized as a major component with an apparent molecular size of 17kDa in TMBZ-staining analysis of the membrane preparation and showed characteristic $\alpha$ (552.5 nm), $\beta$ (522nm), and Soret absorption (416 nm) peaks of a typical reduced c-type cytochrome in the partially purified sample. The internal 43 amino acid sequence of the electron donor was obtained by chemical agent and protease treatments followed by N-terminal sequencing of the resulting fragments. The internal sequence carries lots of lysine residues and a Cys-X-X-Cys-His sequence motif which are the characteristics of typical c-type cytochromes. The analysis of the sequence by FAST or FASTA program, however, did not show any significant similarity to other known heme proteins.

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Standardized microwave extract of Sappan Lignum exerts anti-inflammatory effects through inhibition of NF-κB activation via regulation of heme oxygenase-1 expression

  • Md Anisuzzaman Chowdhury;Moonbum Choi;Wonmin Ko;Hwan Lee;Sam Cheol Kim;Hyuncheol Oh;Eun‑Rhan Woo;Youn‑Chul Kim;Dong‑Sung Lee
    • Molecular Medicine Reports
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    • v.19 no.3
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    • pp.1809-1816
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    • 2019
  • The extract of Sappan Lignum, the heartwood of Caesalpinia sappan L., has been used in medicine to improve blood circulation. Recently, the application of microwave extraction methods has been a major focus of research into the extraction of components from natural sources. In this experiment, we compared the anti-inflammatory effects of Sappan Lignum prepared by heat-70% EtOH extraction (CSE-H-70E) and microwave-70% EtOH extraction (CSE-MW-70E). High-performance liquid chromatography analysis was used to identify the compounds in these extracts. The heat-70% EtOH and microwave-70% EtOH extracts of Sappan Lignum had different chromatograms. CSE-MW-70E significantly inhibited the protein expression of iNOS and COX-2, PGE2, TNF-α, and reduced NO and IL-1β production in macrophages exposed to LPS, whereas, only high concentrations of CSE-H-70E (20 ㎍/ml) resulted in any effects. Furthermore, CSE-MW-70E upregulated heme oxygenase-1 (HO-1) expression. In addition, the use of tin protoporphyrin, an inhibitor of HO-1, confirmed the inhibitory effects of CSE-MW-70E on pro-inflammatory mediators. These results suggested that the CSE-MW-70E-mediated upregulation of HO-1 played an important role in the anti-inflammatory effects of macrophages. Therefore, these findings showed that microwave extraction can be utilized to improve the extraction efficiency and biological activity of Sappan Lignum.

Effects of Washing and Desinewing Treatments on the Composition and Quality Characteristics of Spent Layer Meat (세척 및 결체조직 제거 처리가 노계육의 성분 및 품질 특성에 미치는 영향)

  • Yi, Song-Sop;Mast, Morris G.
    • Korean Journal of Food Science and Technology
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    • v.23 no.6
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    • pp.702-710
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    • 1991
  • The effects of washing and desinewing on the composition, fuctional properties, storage stability and texture of spent layer meat were investigated. Spent layer meat subjected to treatments involving water washing, bicarbonate washing and desinewing showed increases in moisture content and decreases in protein content, hydroxyproline content and heme pigment content. Increases in salt extractable protein content and water holding capacity and decreases in buffering capacity and emulsifying capacity were also observed. The 2-thiobarbituric acid values of washed and desinewed samples increased slowly during storage indicated the increase in storage stability compared to the control. Sample rolls prepared from bicarbonate washed and desinewed thigh and drumstick meats were scored by trained sensory panelists as less tough than the products made of ground thigh and drumstick meats.

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Isoegomaketone Upregulates Heme Oxygenase-1 in RAW264.7 Cells via ROS/p38 MAPK/Nrf2 Pathway

  • Jin, Chang Hyun;So, Yang Kang;Han, Sung Nim;Kim, Jin-Baek
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.510-516
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    • 2016
  • Isoegomaketone (IK) was isolated from Perilla frutescens, which has been widely used as a food in Asian cuisine, and evaluated for its biological activity. We have already confirmed that IK induced the HO-1 expression via Nrf2 activation in RAW264.7 cells. In this study, we investigated the effect of IK on the mechanism of HO-1 expression. IK upregulated HO-1 mRNA and protein expression in a dose dependent manner. The level of HO-1 mRNA peaked at 4 h after $15{\mu}M$ IK treatment. To investigate the mechanisms of HO-1 expression modulation by IK, we used pharmacological inhibitors for the protein kinase C (PKC) family, PI3K, and p38 MAPK. IK-induced HO-1 mRNA expression was only suppressed by SB203580, a specific inhibitor of p38 MAPK. ROS scavengers (N-acetyl-L-cysteine, NAC, and glutathione, GSH) also blocked the IK-induced ROS production and HO-1 expression. Furthermore, both NAC and SB203580 suppressed the IK-induced Nrf2 activation. In addition, ROS scavengers suppressed other oxidative enzymes such as catalase (CAT), glutathione S-transferase (GST), and NADH quinone oxidoreductase (NQO-1) in IK-treated RAW264.7 cells. Taken together, it can be concluded that IK induced the HO-1 expression through the ROS/p38 MAPK/Nrf2 pathway in RAW264.7 cells.

Effects of Polysaccharide Ginsan from Panax ginseng on Liver Function

  • Song, Jie-Young;Medea-Akhalaia;Alexander-Platonov;Kim, Hyung-Doo;Jung, In-Sung;Han, Young-Soo;Yun, Yeon-Sook
    • Archives of Pharmacal Research
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    • v.27 no.5
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    • pp.531-538
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    • 2004
  • Ginsan, a polysaccharide isolated from Panax ginseng, has been shown to be a potent immunomodulator, producing a variety of cytokines such as TNF-a, IL-1$\beta$, IL-2, IL-6, IL-12, IFN-${\gamma}$ and GM-CSF, and stimulating lymphoid cells to proliferate. In the present study, we analyzed some immune functions 1$^{st}$-5$^{th}$ days after ginsan i.p. injection, including the level of non-protein thiols (NPSH) as antioxidants, heme oxygenase (HO) activity as a marker of oxidative stress, zoxazolamine-induced paralysis time and level of hepatic cytochrome P-450 (CYP450) as indices of drug metabolism system, and activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin, and albumin level as indicators of hepatotoxicity. Ginsan in the dose of 100 mg/kg caused marked elevation (1.7-2 fold) of HO activity, decrease of total CYP450 level (by 20-34%), and prolongation of zoxazolamine-induced paralysis time (by 65-70%), and showed some differences between male and female mice. Ginsan treatment did not seem to cause hepatic injury, since serum AST, ALT, and ALP activities and levels of total bilirubin and albumin were not changed.d.

Protective effect of wild ginseng cambial meristematic cells on ᴅ-galactosamine-induced hepatotoxicity in rats

  • Kim, Seok-Joo;Choi, Hyo-Sun;Cho, Hong-Ik;Jin, Young-Woo;Lee, Eun-Kyong;Ahn, Jeung Youb;Lee, Sun-Mee
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.376-383
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    • 2015
  • Background: Panax ginseng has a wide range of biological activities including anti-inflammatory, antioxidant, and immunomodulatory functions. Wild ginseng cambial meristematic cells (CMCs) were obtained from P. ginseng cambium. This study examined the protective mechanism of wild ginseng CMCs against $\small{D}$-galactosamine (GalN)-induced liver injury. GalN, a well-known hepatotoxicant, causes severe hepatocellular inflammatory damage and clinical features similar to those of human viral hepatitis in experimental animals. Methods: Hepatotoxicity was induced in rats using GalN (700 mg/kg, i.p.). Wild ginseng CMCs was administered orally once a day for 2 wks, and then 2 h prior to and 6 h after GalN injection. Results: Wild ginseng CMCs attenuated the increase in serum aminotransferase activity that occurs 24 h after GalN injection. Wild ginseng CMCs also attenuated the GalN-induced increase in serum tumor necrosis factor-${\alpha}$, interleukin-6 level, and hepatic cyclooxygenase-2 protein and mRNA expression. Wild ginseng CMCs augmented the increase in serum interleukin -10 and hepatic heme oxygenase-1 protein and mRNA expression that was induced by GalN, inhibited the increase in the nuclear level of nuclear factor-kappa B, and enhanced the increase in NF-E2-related factor 2. Conclusion: Our findings suggest that wild ginseng CMCs protects liver against GalN-induced inflammation by suppressing proinflammatory mediators and enhancing production of anti-inflammatory mediators.

Studies on Acetanilide p-Hydroxylase in Streptomyces spp. (Streptomyces 속 중의 Acetanilide p-Hydroxylase에 관한 연구)

  • Kim, Jung-Ae;Lee, Sang-Sup
    • YAKHAK HOEJI
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    • v.32 no.5
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    • pp.295-303
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    • 1988
  • For microbial production of acetaminophen, a popular analgesic-antipyretic from aniline, we screened various fungi and bacteria. And we succeeded to some extents in acetaminophen production by successful protoplast fusion between S. lividans and S. globisporus and also between S. rimosus and S. aureofaciens. However, more fertile results might be brought via performing the cloning of acetanilide p-hydroxylation genes of Streptomyces in yeast. This study was initiated to determine whether the acetanilide p-hydroxylase of Streptomyces is cytochrome P-450 species or non-heme iron protein species. The p-hydroxylationactivity on acetanilide in S. aureofaciens ATCC 10762 was found to be unstable on exposing to the air. However, 100,000xg supernatant of the cell free extracts which were prepared in $N_2$ atmosphere showed the p-hydroxylation activity. Characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was not observed in the region of 450nm. Moreover, metyrapone and 2, 6-dichloroindophenol did not affect this enzyme activity, but sodium azide, sodium cyanide, cupric sulfate, cadmium chloride, ${\alpha}$, ${\alpha}'-dipyridyl$, and o-phenanthroline reduced p-hydroxylase activity considerably. S. fradiae NRRL 2702 was shown to have strong p-hydroxylation activity in intact cells. This activity disappeared in its cell free extracts. In its 100,000xg supernatant, however, characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was observed at 446nm. Thus, the results herein presented suggest that acetanilide p-hydroxylase of Streptomyces aureofaciens is not related to cytochrome P-450 and may include non-heme iron protein for its activity. However, it is not clear whether acetanilide p-hydroxylase in S. fradiae belongs to the same category of S. aureofaciens.

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Sparassis crispa (Wulf.) Extract Inhibits IL-1β Stimulated Inflammatory Mediators Production on SW1353 Human Chondrocytes (인간 유래 연골세포에서 꽃송이버섯 추출물의 염증성 매개인자 억제 효과)

  • Kim, Eun-Nam;Jeong, Gil-Saeng
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.305-311
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    • 2018
  • Osteoarthritis (OA) is the most common form of joint disease, characterized by articular cartilage, osteonecrosis, and osteochondral bone erosion. It is an early, progressive disease that combines joint stiffness and joint pain and reduces cartilage function and condition. Interleukin-1 beta ($IL-1{\beta}$) is thought to be important to the pathogenesis of OA and significantly increases the expression of matrix metalloproteinases (MMPs), which play an important role in cartilage degradation in OA. Sparassis crispa (Wulf.) is an edible / medicinal mushroom that has been reported to variety of biological activities. In this study, investigated the Anti-inflammatory effect of Sparassis crispa (Wulf.) ethanol extract (SCE) on $IL-1{\beta}$ stimulated SW1353 chondrocytes. SCE decreased the expression and activity of MMPs by $IL-1{\beta}$ and decreased the levels of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) associated with the inhibition of prostaglandin E2($PGE_2$) in $IL-1{\beta}$ stimulated SW-1353 chondrocytes. In addition, SCE inhibits the expression of MAPK (mitogen-activated protein kinase) and $NF-{\kappa}B$ (nuclear factor-kappa B) signaling in $IL-1{\beta}$ stimulated SW-1353 cells, and SCE inhibits the production of reactive oxygen species (ROS) through heme oxygenase-1 (HO-1) expression. Thus, it is suggested that SCE has a potential as an anti-inflammatory agent in osteoarthritis treatments.

Iron Status and Its Relations with Nutrient Intake, Coffee Drinking, and Smoking in Korean Urban Adults

  • Lee, Joung-Won;Hyun, Wha-Jin;Kwak, Chung-Shil
    • Journal of Community Nutrition
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    • v.5 no.1
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    • pp.44-50
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    • 2003
  • The iron status and its relations with nutrient intake, coffee drinking, and cigarette smoking were evaluated through the blood analysis and 3-day dietary recalls in 102 apparently healthy Korean adults (48 males, 54 menstruating females) aged 20-49 years and living in Daejeon City. Mean values of hemoglobin (Hb) in males and females were 15.5g/dL and 13.2g/dL, mean corpscular hemoglobin concentration (MCHC) 36.0% and 36.8 %, serum iron (SI) 135 $\mu\textrm{g}$/dL and 97 $\mu\textrm{g}$/dL, transferrin saturation (TS) 39.4% and 29.2%, and serum ferritin (Ft) 88.1 $\mu\textrm{g}$/L and 23.4 $\mu\textrm{g}$/L, respectively. For males the prevalences of abnormal values of iron status indicators were 4.2% in Hb, 2.1% in TS, and 4.2% in Ft, and for females 16.7% in Hb, 25.9% in TS, and 35.2% in Ft. Among females 9.3% had abnormal Ft, TS, and Hb, which was considered as iron-defeciency anemia, and 14.8% had abnormal Ft and TS. As a whole, the impaired iron status prevalences were estimated to be 2.1 - 4.2% for males and 9.3 - 35.2% for females. Mean daily intakes of iron and heme-iron were 13.7mg and 1.51mg in males, and 12.3mg and 1.45mg in females. Ft was positively correlated with dietary energy, protein, iron, and vitamin A, Hb with energy and iron, and MCHC with iron and heme iron. Vitamin A also tended to show positive correlations with Hb, SI, and TS. Coffee drinkers taking 3 cups per day or more had higher levels of Hb, MCHC, and Ft in males and MCHC in females, compared to non-coffee drinkers. Higher levels of Hb and MCHC were found in male smokers than in non-smokers. Coffee drinkers took more energy and vitamin A in males and MPF protein in females than non-coffee drinkers. From the above results, it was suggested that the iron status of men was much better than that of women, and the intakes of energy, iron, heme iron, and especially vitamin A were positively associated with the iron status. Cigarette smoking elevated Hb and MCHC, but the effect of coffee drinking on iron staus was not clear. (J Community Nutrition 5(1) : 44∼50, 2003)

Involvement of Peroxynitrite in NO Donor-Induced HO-1 Expression in Rat Articular Chondrocytes (흰쥐 관절연골세포에서 NO donor에 의해 유도된 HO-1 발현에서 peroxynitrite의 관련성 연구)

  • Song, Ju-Dong;Kim, Kang-Mi;Kim, Jong-Min;Yoo, Young-Hyun;Park, Young-Chul
    • Journal of Life Science
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    • v.21 no.4
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    • pp.486-493
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    • 2011
  • Nitric oxide (NO) donors are a potent inducer of heme oxygenase-1 (HO-1). However, it is unclear whether or not HO-1 expression induced by NO donors is a direct consequence of NO released by NO donors. Here, we investigated the effects of NO donors on the expression of HO-1 in primary rat articular chondrocytes. NO donors (SIN-1, SNAP, and SNP) significantly induced the accumulation of HO-1 protein accompanied by an increase in HO-1 mRNA. NO donor-induced HO-1 expression exerted cytoprotection against NO and/or superoxide-induced cell death. Guanylate cyclase signaling was not associated with Nrf2 and HO-1 expression in NO donor-treated chondrocytes. Interestingly, NO scavenger carboxy-PTIO and SOD mimetic TEMPOL markedly inhibited NO donor-induced HO-1 expression in chondrocytes. In addition, NO donor-induced HO-1 expression was completely abrogated by the peroxynitrite scavenger MnTBAP. Since peroxynitrite can be physiologcally formed in the cell through reaction of NO with superoxide, we analyzed whether or not peroxynitrite could directly induce HO-1 expression in chondrocytes. Peroxynitrite treatment in chondrocytes evoked doseand time-dependent Nrf2 and HO-1 expression. These results indicate that HO-1 expression induced by NO donors in rat articular chondrocytes is due to NO-mediated peroxynitrite rather than NO.