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21.
We have previously shown that largemouth bass (Micropterus salmoides) has a remarkable ability to conjugate 4-hydroxy-2-nonenal (4HNE), a mutagenic and cytotoxic α,β-unsaturated aldehyde produced during the peroxidation of lipids. In addition, we have isolated a glutathione S-transferase cDNA (bass GSTA) that encodes a recombinant protein which is highly active in 4HNE conjugation and structurally similar to plaice (Pleuronectes platessa) GSTA. In the present study, HPLC-GST subunit analysis revealed the presence of at least two major GST isoforms in bass liver, with one peak constituting 80% of the total bass liver GST protein. Liquid chromatography mass spectrometry (LC–MS) and electrospray ionization analysis of the major bass GST subunit yielded a molecular weight of 26,396 kDa. Endo-proteinase Lys-C digestion and Edman degradation protein sequencing of this GST peak demonstrated that this protein was encoded by bass GSTA. Analysis of genomic DNA fragments isolated by nested PCR indicated the presence of a GST gene cluster in bass liver that contained GSTA, and was similar to a GST gene cluster characterized by Leaver et al., in plaice. Collectively, our data indicates the presence of a major GST in bass liver involved in the protection against oxidative stress. This GST is part of a gene cluster that may be conserved in certain freshwater and marine fish.  相似文献   
22.
The purpose of the present study was to elucidate in vitro effects of Hg2+, Zn2+, Ni2+ and Cd2+ on cytochrome P4501A1 (CYP1A1) dependent EROD activities in leaping mullet liver microsomes. Fish captured from the most polluted part of Izmir Bay, had highly elevated EROD activities, and induced CYP1A1 protein levels as determined by Western blotting. Although all of the metal ions caused inhibition of the initial velocity of the reaction, Hg2+ and Cd2+ exhibited much higher inhibitory effect at lower concentrations and they were evidently more potent inhibitors than others. The inhibitor concentration giving 50% inhibition (IC50 values) of Zn2+, Ni2+, Cd2+ and Hg2+ of initial EROD activity were 107, 16, 1.3 and 0.15 micromolar, respectively. Glutathione (GSH) at 0.5 mM final concentration, completely reversed Ni2+ and Cd2+ inhibition of EROD activity indicating the protective action of GSH.  相似文献   
23.
Glutathione S-transferases (GSTs) are a class of enzymes that facilitate the detoxification of xenobiotics, and also play important roles in antioxidant defense. We identified two glutathione S-transferase isoforms (VpGSTS, sigma GST; VpGSTO, omega GST) from Venerupis philippinarum by RACE approaches. The open reading frames of VpGSTS and VpGSTO were of 612 bp and 729 bp, encoding 203 and 242 amino acids with an estimated molecular mass of 22.88 and 27.94 kDa, respectively. The expression profiles of VpGSTS and VpGSTO responded to heavy metals and benzo[a]pyrene (B[a]P) exposure were investigated by quantitative real-time RT-PCR. The expression of VpGSTS and VpGSTO were both rapidly up-regulated, however, they showed differential expression patterns to different toxicants. Cd displayed stronger induction of VpGSTS expression with an approximately 12-fold increase than that of VpGSTO with a maximum 6.4-fold rise. Cu exposure resulted in similar expression patterns for both VpGSTS and VpGSTO. For B[a]P exposure, the maximum induction of VpGSTO was approximately two times higher than that of VpGSTS. Altogether, these findings implied the involvement of VpGSTS and VpGSTO in host antioxidant responses, and highlighted their potential as a biomarker to Cd and B[a]P exposure.  相似文献   
24.
Alterations in the glutathione antioxidant system and lipid peroxidation in Avicennia marina were studied under laboratory and field conditions. The activity of glutathione peroxidase (GPx) was found to respond to Zn exposure, and a significant positive relationship between leaf Zn concentration and GPx activity was observed after 96 h and 8 weeks. Lipid hydroperoxides increased proportionally with increasing leaf Zn concentration after 2 and 8 weeks, while no changes in total glutathione were observed. Induction of GPx at 96 h predicted effects at the individual level at a later time interval (reduced biomass at 8 weeks). Results from the field revealed that increasing leaf metal concentration (Zn, Cu or Pb) produced a proportional increase in GPx activity whereas lipid hydroperoxides and total glutathione were not affected. The utility of GPx as an early warning biomarker is suggested, since GPx activity increases in a dose-dependant fashion in response to accumulated leaf metals, and is predictive of later effects on growth.  相似文献   
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26.
以南极硅藻GJ01(Berkeleya rutilans)为对象,分别采用紫外、可见和荧光分光光度法,研究了营养条件等因子对GJ01生长和胞内谷胱甘肽含量的影响,并对培养的GJ01细胞合成谷胱甘肽的条件进行了优化。结果表明,在单因子影响条件下,1.2g/L的碳酸氢钠对南极硅藻GJ01的生长最好,且最有利于谷胱甘肽的积累;0.8mmol/L氯化钙对GJ01的生长最为有利,而0.2mmol/L氯化钙对谷胱甘肽的产生是适宜的;维生素B_(12)和生物素对GJ01胞内谷胱甘肽的产生影响较小。正交试验表明,GJ01细胞产生谷胱甘肽的最优条件是:光照周期为16h/8h、NaHCO_3为1.6g/L、Ca~(2 )为0.4mmol/L、Cys为10mmol/L。该结果为利用南极硅藻GJ01生产谷胱甘肽提供了参考和依据。  相似文献   
27.
The health of red spruce is adversely affected by exposure to acid fog. Epicuticular wax structure over epistomatal chambers, mesophyll membrane calcium and changes in redox state of glutathione are affected in both chamber and field studies and are correlated with leaf exposure to acid fog, but not with soil or total leaf calcium concentrations. Experiments tracking fog deposition under conditions of illumination or darkness, with or without a metabolic inhibitor, support the mechanism of diffusiophoresis linked with photosynthetic gas exchange. Based on deposition pattern and leaf morphology, a stomatal pathway for water and hydrogen ion penetration to leaf interior is proposed. Ecophysiological parameters are discussed in relation to differential sensitivities of red spruce and white pine to ozone and acid fog.  相似文献   
28.
Glutathione S-transferase (GST) and heat shock proteins (hsps) 40, 60, 70 and 90 were determined by immunoblotting using actin as an internal control in Mytilus edulis from one station outside (site1) and three stations within (sites 2-4) Cork Harbour, Ireland. Comparisons were made between gill and digestive gland and between sites. Gill shows generally higher hsp 60, 70 and 90 while digestive gland has higher hsp 40. Site 1 showed higher gill hsps 40 and 70 than sites 2-4 while gill GST was higher in sites 3 and 4 than 1 and 2. Comparison with sites in the North Sea (site 5: outside Tj?rn? in The Koster archipelago in the Skagerack) and Baltic Sea (site 6: Ask? island) also revealed lower hsps 40 and 70 in site 6 (low salinity) than site 5 (high salinity) although hsps 60, 70 and 90 were detectable in digestive gland unlike sites 1-4. Previously, only hsp 70 had been studied at these sites [Mar. Environ. Res. 39. (1995), 181]. At the mRNA level, gill hsp 70 is 80-fold higher at Tj?rn? than Ask?. These data suggest that, while salinity may slightly decrease hsp 40 and 70, both hsp 70 and GST are selectively up-regulated by approx. 10- and 3-fold, respectively, at Tj?rn? compared to the other sites which we attribute to exposure to more widely fluctuating pollution levels.  相似文献   
29.
用酶标仪采用DTNB比色法测定栉孔扇贝(Chlamys farreri)血淋巴中谷胱甘肽还原酶(GR)的活性,并研究pH值、温度、激活剂和抑制剂对血清中该酶活性的影响及酶的热稳定性。结果表明,栉孔扇贝血细胞中GR比活力明显高于血清。血清中GR的最适pH值因缓冲液的不同而稍有差异:用PBS为缓冲液时,最适pH值为7.0;Tris-HCl为缓冲液时,最适pH值为7.5。酶的最适温度为60℃。将血清在20℃,30℃,40℃,50℃,60℃,70℃和80℃温度的水浴中放置30min后于20℃,pH7.5条件下测定该酶的热稳定性,结果显示50℃时酶的比活力最高,低于或高于此温度的酶比活力均下降,80℃时酶活性完全丧失。栉孔扇贝血淋巴中GR属中温型中性酶类,低温下较稳定。EDTA对该酶活性有促进作用,硫酸铜有抑制作用。  相似文献   
30.
苯并(a)芘和芘对梭鱼肝脏谷胱甘肽过氧化酶活性的影响   总被引:5,自引:0,他引:5  
在实验生态条件下,浓度范围0.1-50μg/L的苯并(a)芘和芘的短期暴露造成梭鱼肝脏GPx活性先抑制后诱导的效应,相同的污染物,高浓度组对GPx活性的抑制大于低浓度组,出现GPx活性诱导的时间早于低浓度组;相同浓度下,苯并(a)芘对GPx活的作用大于芘,这间接反映了苯并(a)芘和芘的毒性大小,这些结果说明梭鱼肝脏GPx活性与苯并(a)芘和芘暴露有一定的相关性,可以作为海洋环境污染监测的一种生物标志物。  相似文献   
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