Glutathione-S-transferase

谷胱甘肽 - S - 转移酶
  • 文章类型: Journal Article
    在目前的研究中,评估了两种商业工业大麻(IH)纤维品种(V1:CFX-2和V2:Henola)调节盐诱导的氧化应激代谢的能力。30天,在温室环境中使用五种不同的盐度处理(0、50、80、100、150和200mMNaCl)栽培植物。过氧化氢(H2O2),丙二醛(MDA),和脂氧合酶(LOX)和抗氧化酶(超氧化物歧化酶(SOD),过氧化氢酶,愈创木酚过氧化物酶(GPOD),抗坏血酸过氧化物酶(APX),谷胱甘肽还原酶(GR),和谷胱甘肽-S-转移酶(GST))在完全扩张的叶片中进行了评估。在200和100mMNaCl浓度下,分别,生理盐水治疗后30天,V1和V2中的植物没有存活。在80mMNaCl,V2的叶片显示出更高的H2O2,MDA,和LOX比V1。较高的SOD,CAT,GPOD,APX,GR,V1至100mMNaCl的叶片中的GST活性导致H2O2和MDA水平降低。在80mMNaCl,V2证明了抗氧化剂防御机制的完全失败。这些结果表明,V1比V2表现出更强的耐盐性,部分原因是抗氧化代谢更好。
    In the current study, two commercial industrial hemp (IH) fiber varieties (V1: CFX-2 and V2: Henola) were assessed for their ability to regulate salt-induced oxidative stress metabolism. For 30 days, plants were cultivated in greenhouse environments with five different salinity treatments (0, 50, 80, 100, 150, and 200 mM NaCl). Hydrogen peroxide (H2O2), malondialdehyde (MDA), and lipoxygenase (LOX) and antioxidant enzymes (superoxide dismutase (SOD), catalase, guaiacol peroxidase (GPOD), ascorbate peroxidase (APX), glutathione reductase (GR), and glutathione-S-transferase (GST)) were assessed in fully expanded leaves. At 200 and 100 mM NaCl concentrations, respectively, 30 days after saline treatment, plants in V1 and V2 did not survive. At 80 mM NaCl, the leaves of V2 showed higher concentrations of H2O2, MDA, and LOX than those of V1. Higher SOD, CAT, GPOD, APX, GR, and GST activity in the leaves of V1 up to 100 mM NaCl resulted in lower levels of H2O2 and MDA. At 80 mM NaCl, V2 demonstrated the total failure of the antioxidant defense mechanism. These results reveal that V1 demonstrated stronger salt tolerance than V2, in part due to better antioxidant metabolism.
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  • 文章类型: Journal Article
    尼姆,一种生物农药,为桑树种植中常用的合成杀虫剂提供了更安全的替代品,会伤害蚕。本研究旨在研究泰国印度籽提取物对泰国多伏杂交家蚕所有龄幼虫的影响。BombyxmoriL.,DokBua菌株,关注死亡率和酯酶(EST)和谷胱甘肽S-转移酶(GST)酶的活性。使用浸叶法评估急性毒性。结果表明,年轻的幼龄婴儿的死亡率往往高于老年婴儿。第一龄幼虫的死亡率最高,为94%,而LC50在72h时在5.23mgL-1的第三龄中最高。这种趋势与EST和GST的活性一致,在第一龄幼虫的全身和第五龄幼虫的中肠组织中进行了评估。随着提取物浓度的增加,第一龄和第五龄幼虫的EST活性降低,而GST活性增加。这些发现突出表明,印em提取物对所有幼龄幼虫都是有毒的,商品及服务税在排毒中起着至关重要的作用,特别是在泰国多伏杂交蚕的全身。
    Neem, a biopesticide, offers a safer alternative to the synthetic insecticides commonly used in mulberry cultivation, which can harm silkworms. This study aimed to investigate the effects of Thai neem seed extract on all instar larvae of the Thai polyvoltine hybrid silkworm, Bombyx mori L., Dok Bua strains, focusing on the mortality rate and the activities of esterase (EST) and glutathione S-transferases (GST) enzymes. Acute toxicity was assessed using the leaf-dipping method. Results showed that the mortality rate tended to be higher in younger instars than in older ones. The first instar larvae exhibited the highest mortality rate at 94%, whereas the LC50 was highest in the third instar at 5.23 mg L-1 at 72 h. This trend aligns with the activities of EST and GST, which were evaluated in the whole bodies of the first instar larvae and the midgut tissue of fifth instar larvae. As the extract concentration increased, EST activity decreased while GST activity increased in both the first and fifth instar larvae. These findings highlight that neem extract is toxic to all instar larvae, with GST playing a crucial role in detoxification, particularly in the whole body of the Thai polyvoltine hybrid silkworm.
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  • 文章类型: Journal Article
    男性生殖系统可能容易受到空气污染等环境污染物的影响,因此有必要对男性生殖功能失调的潜在机制进行彻底调查。本研究旨在研究Wistar大鼠模型中柴油废气的过滤部分(主要包含气体)对雄性生殖功能的影响。成年雄性大鼠随机分为三组(n=8/组):对照组(未暴露)组(CG-A),WBE舱内的清洁空气组(CAG-A),WBE舱室(FDG-A)中过滤的柴油机排气组。CAG-A和FDG-A的暴露方案为6h/天×5d/周×6周,评价精子参数,睾丸组织病理学,激素定量(睾酮,LH,FSH,17β-雌二醇,和催乳素),和GST水平进行。结果表明,WBE到FDE导致精子浓度显着下降(p=0.008,CG-AvsFDG-A;p=0.014,CAG-AvsFDG-A),运动性(p=0.008,CG-A对FDG-A;p=0.029,CAG-A对FDG-A),血清睾酮(p=0.024,CG-AvsFDG-A;p=0.007,CAG-AvsFDG-A),睾丸睾酮(p=0.008,CG-AvsFDG-A;p=0.028,CAG-AvsFDG-A),17β-雌二醇(p=0.007,CG-A与FDG-A),和GST水平(p=0.0002,CG-AvsFDG-A;p=0.0019,CAG-AvsFDG-A)。这些发现表明,在暴露于FDE的成年大鼠中,睾丸内环境中睾丸激素-雌二醇平衡的破坏没有其他主要垂体激素的显着改变。由于氧化失衡,FDE中主要存在的气体成分会导致睾丸损伤。这强调了FDE暴露和男性生殖结果受损的因果关系。
    The possible vulnerability of the male reproductive system to environmental pollutants such as air pollution necessitates a thorough investigation of the underlying mechanisms involved in the dysregulation of male reproductive function. The present study was designed to investigate the influence of the filtered fraction of diesel exhaust (predominantly comprising gases) on male reproductive function in Wistar rat model. Adult male rats were randomly assigned into three groups (n=8/group): Control (unexposed) group (CG-A), the Clean air group in WBE chamber (CAG-A), and Filtered diesel exhaust group in WBE chamber (FDG-A). The exposure protocol for CAG-A and FDG-A was 6 h/day x 5d/week x 6 weeks,evaluation of sperm parameters, testicular histopathology, quantification of hormones (testosterone, LH, FSH, 17β-Estradiol, and prolactin), and GST levels were performed. Results showed that WBE to FDE leads to a significant decline in sperm concentration (p=0.008, CG-A vs FDG-A; p=0.014, CAG-A vs FDG-A), motility (p=0.008, CG-A vs FDG-A; p=0.029, CAG-A vs FDG-A), serum testosterone (p=0.024, CG-A vs FDG-A; p=0.007, CAG-A vs FDG-A), testicular testosterone (p=0.008, CG-A vs FDG-A; p=0.028, CAG-A vs FDG-A), 17β-Estradiol (p=0.007, CG-A vs FDG-A), and GST levels (p=0.0002, CG-A vs FDG-A; p=0.0019, CAG-A vs FDG-A). These findings demonstrate the disruption of testosterone-estradiol balance in the intratesticular milieu without significant alterations in other principal pituitary hormones in adult rats exposed to FDE. The predominant presence of gaseous components in FDE can cause testicular damage due to oxidative imbalance. This underscores the causality of FDE exposure and impaired male reproductive outcomes.
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  • 文章类型: English Abstract
    OBJECTIVE: To search for possible connections between the anti-inflammatory activity of monocytes (PAM) and the activity of glutathione metabolic enzymes: glutathione reductase (GR) and glutathione-S-transferase (GT) in patients with depressive states (DS) within various mental pathologies, as well as between the studied biological parameters and clinical condition of patients.
    METHODS: Sixty-one women, aged 18-56 years, with DC were examined before and after treatment. Symptom severity was assessed using the Positive and Negative Syndrome Scale (PANSS) and the Hamilton Depressive Symptom Rating Scale (HDRS-21). The control group included 23 women of the corresponding age without mental pathology. Biological parameters were assessed in the peripheral blood of patients and healthy people.
    RESULTS: Patients with a high level of PAM compared to the control (p<0.001) (subgroup 1, n=31) and with a low (at the control level) level (subgroup 2, n=30) were identified. In the subgroup 1, the values of GR and GT were significantly lower than in patients of subgroup 2 (p<0.05 and p<0.01, respectively). Negative correlations between the level of PAM before treatment and GR before and after treatment were revealed in patients who responded to treatment (r=-0.67; p=0.0041; r=-0.76; p=0.0001).
    CONCLUSIONS: The results may indicate the inverse relationship between the level of PAM and the activity of GR and GT, which are involved in the pathogenesis of DC, and can also serve as criteria for assessing the response of patients to treatment.
    UNASSIGNED: Поиск возможных связей между противовоспалительной активностью моноцитов (ПАМ) и активностью ферментов метаболизма глутатиона: глутатионредуктазы (ГР) и глутатион-S-трансферазы (ГТ) у пациентов с депрессивными состояниями (ДС) в рамках различной психической патологии, а также между изученными биологическими параметрами и клиническим состоянием больных.
    UNASSIGNED: Обследовали до и после лечения 61 женщину в возрасте 18—56 лет с ДС. Выраженность симптомов оценивали в баллах по шкале позитивных и негативных синдромов (PANSS) и шкале Гамильтона для оценки тяжести депрессивных симптомов (HDRS-21). Контрольная группа включала 23 женщины соответствующего возраста без психической патологии. Биологические параметры определяли в периферической крови больных и здоровых.
    UNASSIGNED: Выделены больные с высоким, по сравнению с контролем (p<0,001), уровнем ПАМ (подгруппа 1, n=31) и с низким (на уровне контроля) его значением (подгруппа 2, n=30). В подгруппе 1 значения ГР и ГТ были значительно ниже их значений у больных в подгруппе 2 (p<0,05 и p<0,01 соответственно). У больных, отвечавших на лечение, выявлены отрицательные корреляции между уровнем ПАМ до лечения и ГР до и после лечения (r=–0,67; p=0,0041 и r=–0,76; p<0,0001).
    UNASSIGNED: Полученные результаты могут свидетельствовать о существовании обратной связи между уровнем ПАМ и активностью ГР и ГТ и возможном участии выявленных молекулярных механизмов в развитии системного иммунного воспаления в патогенезе ДС, а также могут служить в качестве критериев оценки ответа больных на лечение.
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  • 文章类型: Journal Article
    全球变暖,热浪,季节性干旱对农作物构成严重威胁,比如葡萄藤,它们的次级代谢物很有价值,这对葡萄酒行业至关重要。区分开放领域中不同环境因素的影响是具有挑战性的。在本研究中,将长相思的体外培养浆果暴露于单个和组合的胁迫因子中,以研究其对硫醇前体生物合成的影响。我们的结果证实了葡萄积累过程的复杂性和极端反应性。然而,它们还表明热应激对Cys-3SH前体的产生具有积极作用。此外,我们确定了几个候选基因,例如VvGST和VvGGT,它们可能参与生物合成并被一致调节。尽管如此,我们无法最终确定压力对其他前体生物合成的影响,也无法提出有关其调节的假设。
    Global warming, heat waves, and seasonal drought pose serious threats to crops, such as grapevine, that are valued for their secondary metabolites, which are of primary importance for the wine industry. Discriminating the effects of distinct environmental factors in the open field is challenging. In the present study, in vitro cultured berries of Sauvignon Blanc were exposed to individual and combined stress factors to investigate the effects on the biosynthesis of the thiol precursors. Our results confirm the complexity and extreme reactivity of the accumulation process in grapes. However, they also indicate that heat stress has a positive effect on the production of the Cys-3SH precursor. Moreover, we identified several candidate genes, such as VvGSTs and VvGGT that are potentially involved in biosynthesis and consistently modulated. Nonetheless, we were unable to conclusively determine the effects of stresses on the biosynthesis of other precursors nor could we formulate hypotheses regarding their regulation.
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  • 文章类型: Journal Article
    植物含有多种潜在的植物化学物质,这些化学物质是特定于目标的,对人体健康毒性较小。本研究旨在研究肾素的代谢组学概况(L。)肖特及其通过靶向谷胱甘肽-S-转移酶控制蚊子的潜力,专注于对淡色库蚊的杀幼虫活动。粗提物(CEs)使用乙醇制备,乙酸乙酯和正己烷。CEs已用于灭蚊生物测定的评估。通过气相色谱-质谱(GC-MS)对每种CE进行了CE的代谢组学分析。幼虫死亡率最高,LC50最小的最有效的CE是己烷CE。然后,碱性磷酸酶(ALP)活性,在用己烷CE处理的幼虫中评估谷胱甘肽-S-转移酶(GST)活性。结果显示蛋白质含量下降,ALP活性的诱导,和减少GST活性。最后,采用分子对接和动态模拟技术来评估己烷植物化学物质与GST蛋白之间的相互作用。D-(+)-葡糖醛酸,3TMS衍生物和癸二酸,2TMS衍生物对GST蛋白显示出最佳的结合亲和力,表明它们对酶解毒功能的干扰,可能导致杀虫剂代谢能力下降。
    Plants contain a wide range of potential phytochemicals that are target-specific, and less toxic to human health. The present study aims to investigate the metabolomic profile of Nephrolepis exaltata (L.) Schott and its potential for mosquito control by targeting Glutathione-S-Transferase, focusing on the larvicidal activity against Culex pipiens. Crude extracts (CEs) were prepared using ethanol, ethyl acetate and n-hexane. CEs have been used for assessment of mosquitocidal bioassay. The metabolomic analyses for CEs were characterized for each CE by gas chromatography-mass spectrometry (GC-MS). The most efficient CE with the highest larval mortality and the least LC50 was the hexane CE. Then, alkaline phosphatase (ALP) activity, and glutathione-S-transferase (GST) activity were assessed in larvae treated with the hexane CE. The results demonstrated a decline in protein content, induction of ALP activity, and reduction in GST activity. Finally, molecular docking and dynamic simulation techniques were employed to evaluate the interaction between the hexane phytochemicals and the GST protein. D-(+)-Glucuronic acid, 3TMS derivative and Sebacic acid, 2TMS derivative showed best binding affinities to GST protein pointing to their interference with the enzyme detoxification functions, potentially leading to reduced ability to metabolize insecticides.
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  • 文章类型: Journal Article
    褐飞虱(BPH),Nilaparvatalugens是水稻的一种毁灭性农业害虫,他们已经对许多杀虫剂产生了抗药性。在这项研究中,我们评估了BPH若虫对Nitenpyra的反应,吡虫啉,和etofenprox使用接触和饮食生物测定法,并研究了BPH谷胱甘肽-S-转移酶(GST)的潜在功能多样性,羧酸酯酶(CarE)和细胞色素P450单加氧酶(P450)对这些杀虫剂。硝基吡喃对BPH的接触和摄入毒性均显着高于吡虫啉或依托芬普司。在每种杀虫剂的LC50浓度下,他们引发了对商品及服务税的不同反应,CarE,和P450活动,和每种杀虫剂诱导至少一种解毒酶活性。这些杀虫剂几乎抑制了所有测试的GST的表达,CarE,和P450基因在接触生物测定中,但在饮食生物测定中诱导了这些基因的转录水平。NlGSTD2表达的沉默对接触试验中BPH对氮吡喃胺的敏感性和饮食试验中吡虫啉的敏感性影响最大。NlCE沉默后,接触试验中BPH对杀虫剂的敏感性增加最大的是etofenprox,而饮食测试是氮吡仑。敲除NlCYP408A1导致BPH对杀虫剂的敏感性在接触试验中增加最多的是硝普兰,而饮食测试是吡虫啉。一起来看,这些发现表明,NlGSTD2,NlCE,NlCYP408A1在不同类型杀虫剂对BPH的接触和摄入毒性的解毒中起着不可或缺的作用,这对于制定新的吸虫防治策略具有重要意义。
    The brown planthopper (BPH), Nilaparvata lugens is a devastating agricultural pest of rice, and they have developed resistance to many pesticides. In this study, we assessed the response of BPH nymphs to nitenpyram, imidacloprid, and etofenprox using contact and dietary bioassays, and investigated the underlying functional diversities of BPH glutathione-S-transferase (GST), carboxylesterase (CarE) and cytochrome P450 monooxygenase (P450) against these insecticides. Both contact and ingestion toxicity of nitenpyram to BPH were significantly higher than either imidacloprid or etofenprox. Under the LC50 concentration of each insecticide, they triggered a distinct response for GST, CarE, and P450 activities, and each insecticide induced at least one detoxification enzyme activity. These insecticides almost inhibited the expression of all tested GST, CarE, and P450 genes in contact bioassays but induced the transcriptional levels of these genes in dietary bioassays. Silencing of NlGSTD2 expression had the greatest effect on BPH sensitivity to nitenpyram in contact test and imidacloprid in dietary test. The sensitivities of BPH to insecticide increased the most in the contact test was etofenprox after silencing of NlCE, while the dietary test was nitenpyram. Knockdown of NlCYP408A1 resulted in BPH sensitivities to insecticide increasing the most in the contact test was nitenpyram, while the dietary test was imidacloprid. Taken together, these findings reveal that NlGSTD2, NlCE, and NlCYP408A1 play an indispensable role in the detoxification of the contact and ingestion toxicities of different types of insecticides to BPH, which is of great significance for the development of new strategies for the sucking pest control.
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  • 文章类型: Journal Article
    弱球雕塑是Ixodidae家族中的一种蜱,在其首选寄主中具有马匹和水葫芦。在这项研究中,Piperaduncum精油(EO)及其主要成分的杀螨活性,Dillapiole,针对A的幼虫进行评估,以建立致死浓度值并评估这些化合物对蜱酶的影响。Dillapiole对蜱的活性(LC50=3.38mg/mL;95%CI=3.24至3.54)比P.aduncumEO(LC50=3.49mg/mL;95%CI=3.36至3.62)稍高。α-酯酶(α-EST)活性,β-酯酶(β-EST),与所有浓度(LC5,LC25,LC50和LC75)的对照相比,用Dillapiole处理的A.scultum幼虫中的谷胱甘肽-S-转移酶(GST)酶显示出显着增加,使用P.aduncumEO获得了类似的结果,除了α-EST,在最高浓度(LC75)下与对照没有差异。乙酰胆碱酯酶(AChE)活性的结果表明,在两个较低的浓度(LC5和LC25)下,酶活性增加,而在两个较高的浓度下,酶活性降低,与对照相比,致死浓度(LC50和LC75)。这些结果表明了这些天然杀螨剂的潜在作用机制,并可以为潜在的植物衍生制剂的未来开发提供指导。
    Amblyomma sculptum is a species of tick in the family Ixodidae, with equids and capybaras among its preferred hosts. In this study, the acaricidal activity of the essential oil (EO) from Piper aduncum and its main component, Dillapiole, were evaluated against larvae of A. sculptum to establish lethal concentration values and assess the effects of these compounds on tick enzymes. Dillapiole exhibited slightly greater activity (LC50 = 3.38 mg/mL; 95% CI = 3.24 to 3.54) than P. aduncum EO (LC50 = 3.49 mg/mL; 95% CI = 3.36 to 3.62) against ticks. The activities of α-esterase (α-EST), β-esterase (β-EST), and glutathione-S-transferase (GST) enzymes in A. sculptum larvae treated with Dillapiole showed a significant increase compared to the control at all concentrations (LC5, LC25, LC50 and LC75), similar results were obtained with P. aduncum EO, except for α-EST, which did not differ from the control at the highest concentration (LC75). The results of the acetylcholinesterase (AChE) activity show an increase in enzyme activity at the two lower concentrations (LC5 and LC25) and a reduction in activity at the two higher, lethal concentrations (LC50 and LC75) compared to the control. These results suggest potential mechanisms of action for these natural acaricides and can provide guidance for the future development of potential plant-derived formulations.
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  • 文章类型: Journal Article
    谷胱甘肽-S-转移酶(GSTs)是II期解毒系统的重要组成部分,可保护生物体免受外源性物质和有害毒素(例如1-氯-2,4-二硝基苯(CDNB))诱导的氧化应激。在嗜热四膜虫中,TtGSTm34基因先前被报道为对CDNB治疗最敏感的GST基因之一(LD50=0.079mM).本研究旨在确定用CDNB和谷胱甘肽(GSH)重组表达和纯化的TtGSTm34的动力学特征。在将TtGSTm34的660-bp全长ORF克隆到pIGF-1载体中后,在嗜热T.thermophila中重组产生TtGSTm34-8xHis为25kDa蛋白。通过AlphaFold和PyMOL程序构建的TtGSTm34蛋白的三维模型证实其具有结构上保守和折叠的GST结构域。通过SDS-PAGE和Western印迹分析证实了TtGSTm34-8xHis的重组产生。双重亲和层析策略有助于纯化TtGSTm34-8xHis约3166倍。纯化的重组TtGSTm34-8xHis以CDNB(190µmol/min/mg)为底物表现出显着的高酶活性。酶动力学分析显示,以GSH为底物的Km值为0.68mM,以CDNB为底物的Km值为0.40mM,证实其对CDNB的预期高亲和力。确定的最佳pH和温度为7.0和25°C,分别。乙丙炔酸完全抑制TtGSTm34-8xHis酶活性。这些结果表明,嗜热T.thermophila的TtGSTm34在外源性物质的解毒中起主要作用,例如CDNB,作为水生原生生物抵御氧化损伤的第一道防线。
    Glutathione-S-transferase enzymes (GSTs) are essential components of the phase II detoxification system and protect organisms from oxidative stress induced by xenobiotics and harmful toxins such as 1-chloro-2,4-dinitrobenzene (CDNB). In Tetrahymena thermophila, the TtGSTm34 gene was previously reported to be one of the most responsive GST genes to CDNB treatment (LD50 = 0.079 mM). This study aimed to determine the kinetic features of recombinantly expressed and purified TtGSTm34 with CDNB and glutathione (GSH). TtGSTm34-8xHis was recombinantly produced in T. thermophila as a 25-kDa protein after the cloning of the 660-bp full-length ORF of TtGSTm34 into the pIGF-1 vector. A three-dimensional model of the TtGSTm34 protein constructed by the AlphaFold and PyMOL programs confirmed that it has structurally conserved and folded GST domains. The recombinant production of TtGSTm34-8xHis was confirmed by SDS‒PAGE and Western blot analysis. A dual-affinity chromatography strategy helped to purify TtGSTm34-8xHis approximately 3166-fold. The purified recombinant TtGSTm34-8xHis exhibited significantly high enzyme activity with CDNB (190 µmol/min/mg) as substrate. Enzyme kinetic analysis revealed Km values of 0.68 mM with GSH and 0.40 mM with CDNB as substrates, confirming its expected high affinity for CDNB. The optimum pH and temperature were determined to be 7.0 and 25 °C, respectively. Ethacrynic acid inhibited fully TtGSTm34-8xHis enzyme activity. These results imply that TtGSTm34 of T. thermophila plays a major role in the detoxification of xenobiotics, such as CDNB, as a first line of defense in aquatic protists against oxidative damage.
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  • 文章类型: Journal Article
    谷胱甘肽-S-转移酶是异源物质和氧化损伤产物的细胞解毒的关键。GSTs催化谷胱甘肽(GSH)与亲电试剂的反应以形成稳定的硫醚加合物。GSTK1-1是线粒体基质中主要的GST同工型,但是GSTA1-1和GSTA4-4同工型也被认为在线粒体中,它们在转化细胞中的分布发生变化,从而潜在地提供癌症特异性靶标。GST底物1-氯-2,4-二硝基苯(CDNB)的线粒体靶向版本,MitoCDNB,已用于操纵线粒体GSH池。为了巧妙地利用这种方法来针对癌症中的特定GST同工型,在这里,我们已经确定了GSTK1-1,GSTA1-1和GSTA4-4的人亚型相对于GSH和CDNB的kcat/Km.我们展示了GSH和CDNB类似物之间的GST催化反应速率如何通过两个吸电子取代基进行修饰,并通过线粒体靶向三苯基鳞在氯苯环上的位置来调节线粒体靶向底物的活性。现在可以利用这些发现来选择性地破坏表达特定GST同工型的癌细胞的线粒体GSH库。
    Glutathione-S-transferases are key to the cellular detoxification of xenobiotics and products of oxidative damage. GSTs catalyse the reaction of glutathione (GSH) with electrophiles to form stable thioether adducts. GSTK1-1 is the main GST isoform in the mitochondrial matrix, but the GSTA1-1 and GSTA4-4 isoforms are also thought to be in the mitochondria with their distribution altering in transformed cells, thus potentially providing a cancer specific target. A mitochondria-targeted version of the GST substrate 1-chloro-2,4-dinitrobenzene (CDNB), MitoCDNB, has been used to manipulate the mitochondrial GSH pool. To finesse this approach to target particular GST isoforms in the context of cancer, here we have determined the kcat/Km for the human isoforms of GSTK1-1, GSTA1-1 and GSTA4-4 with respect to GSH and CDNB. We show how the rate of the GST-catalysed reaction between GSH and CDNB analogues can be modified by both the electron withdrawing substituents, and by the position of the mitochondria-targeting triphenylphosphonium on the chlorobenzene ring to tune the activity of mitochondria-targeted substrates. These findings can now be exploited to selectively disrupt the mitochondrial GSH pools of cancer cells expressing particular GST isoforms.
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