Monocrotophos

久效磷
  • 文章类型: Journal Article
    有机磷农药(OPPs)是水生生态系统中重要的化学应激源,最近它们吸引了越来越多的关注。然而,不同OPPs对碳循环的影响尚不清楚,特别是那些功能性但无法培养的微生物。这项研究调查了在敌敌畏存在下湖泊水生微生物群落的变化,久效磷,氧乐果和对硫磷。所有OPP均显著抑制生物量(p<0.05)和碳循环相关cbbLG基因表达(p<0.01),改变了水生微生物群落结构,互动,和组装。方差分配分析表明,农药类型对微生物生物量和群落结构的影响更大。其中农药浓度在碳循环中起着更重要的作用。通过Wood-Ljungdahl途径分析cbbLG基因和PICRUSt2,木脂杆菌和麻黄菌科同化的无机碳,而CyanobiumPCC-6307是Calvin-Benson-Bassham循环。这项工作提供了一个更深入的见解的行为和机制的微生物群落变化在水生生物多样性的反应,并明确揭示了OPP对其碳循环功能的影响。
    Organophosphorus pesticides (OPPs) are important chemical stressors in aquatic ecosystems, and they attract increasing more attentions recently. However, the impacts of different OPPs on carbon cycling remain unclear, particularly for those functional-yet-uncultivable microbes. This study investigated the change in lake aquatic microbial communities in the presence of dichlorvos, monocrotophos, omethoate and parathion. All OPPs significantly inhibited biomass (p < 0.05) and the expression of carbon cycle-related cbbLG gene (p < 0.01), and altered aquatic microbial community structure, interaction, and assembly. Variance partitioning analysis showed a stronger impact of pesticide type on microbial biomass and community structure, where pesticide concentration played more significant roles in carbon cycling. From analysis of cbbLG gene and PICRUSt2, Luteolibacter and Verrucomicrobiaceae assimilated inorganic carbon through Wood-Ljungdahl pathway, whereas it was Calvin-Benson-Bassham cycle for Cyanobium PCC-6307. This work provides a deeper insight into the behavior and mechanisms of microbial community change in aquatic system in response to OPPs, and explicitly unravels the impacts of OPPs on their carbon-cycling functions.
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  • 文章类型: Journal Article
    芽孢杆菌属,尤其是在根瘤菌中,以其促进植物生长的能力和减轻几种胁迫条件的有效性而闻名。本研究旨在利用本地蜡样芽孢杆菌PM38降解毒死蜱(CP)等4种有机磷农药(OPs),profenofos(PF),久效磷(MCP),和乐果(DMT),以减轻这些农药对棉花作物生长的不利影响。菌株PM38表现出与其他芽孢杆菌属物种不同的独特特性。这些包括胞外酶的产生,氰化氢,胞外多糖,吲哚-3-乙酸(166.8μg/mL),铁载体(47.3μg/mL),1-氨基环丙烷-1-羧酸脱氨酶活性(32.4μg/mL),和磷溶解(162.9μg/mL),所有在较高浓度下观察到。该菌株还显示出对盐度(1200mM)的耐受性,干旱(20%PEG-6000),以及铜和镉(1200mg/L)。多应激反应基因的扩增,比如acds,ituC,czcD,nifH,SFP,和pqqE,进一步证实了菌株PM38对植物生长的调控和对非生物胁迫的耐受能力。在高效液相色谱(HPLC)分析之后,结果显示与第一个动力学模型具有惊人的兼容性。菌株PM38能有效降解CP(98.4%),PF(99.7%),MCP(100%),和浓度为300ppm的DMT(95.5%),在最佳pH条件下在35°C下在48小时内,显示高的测定系数(R2)分别为0.974、0.967、0.992和0.972。傅里叶变换红外光谱(FTIR)分析,mpd,菌株PM38中的opdA基因进一步支持降解OPs的潜力。此外,在胁迫条件下,用PM38接种棉花幼苗可以改善根长。菌株PM38的接种通过最小化脯氨酸来降低应力,硫代巴比妥酸反应性化合物,和电解质泄漏。菌株PM38有可能成为能够改善全球粮食安全和管理受污染地点的生物害虫防治剂的良好的多重胁迫耐受性选择。
    Bacillus genera, especially among rhizobacteria, are known for their ability to promote plant growth and their effectiveness in alleviating several stress conditions. This study aimed to utilize indigenous Bacillus cereus PM38 to degrade four organophosphate pesticides (OPs) such as chlorpyrifos (CP), profenofos (PF), monocrotophos (MCP), and dimethoate (DMT) to mitigate the adverse effects of these pesticides on cotton crop growth. Strain PM38 exhibited distinct characteristics that set it apart from other Bacillus species. These include the production of extracellular enzymes, hydrogen cyanide, exopolysaccharides, Indol-3-acetic acid (166.8 μg/mL), siderophores (47.3 μg/mL), 1-aminocyclopropane-1-carboxylate deaminase activity (32.4 μg/mL), and phosphorus solubilization (162.9 μg/mL), all observed at higher concentrations. This strain has also shown tolerance to salinity (1200 mM), drought (20% PEG-6000), and copper and cadmium (1200 mg/L). The amplification of multi-stress-responsive genes, such as acdS, ituC, czcD, nifH, sfp, and pqqE, further confirmed the plant growth regulation and abiotic stress tolerance capability in strain PM38. Following the high-performance liquid chromatography (HPLC) analysis, the results showed striking compatibility with the first kinetic model. Strain PM38 efficiently degraded CP (98.4%), PF (99.7%), MCP (100%), and DMT (95.5%) at a concentration of 300 ppm over 48 h at 35 °C under optimum pH conditions, showing high coefficients of determination (R2) of 0.974, 0.967, 0.992, and 0.972, respectively. The Fourier transform infrared spectroscopy (FTIR) analysis and the presence of opd, mpd, and opdA genes in the strain PM38 further supported the potential to degrade OPs. In addition, inoculating cotton seedlings with PM38 improved root length under stressful conditions. Inoculation of strain PM38 reduces stress by minimizing proline, thiobarbituric acid-reactive compounds, and electrolyte leakage. The strain PM38 has the potential to be a good multi-stress-tolerant option for a biological pest control agent capable of improving global food security and managing contaminated sites.
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  • 文章类型: Journal Article
    最近,农药的使用,农田中的除草剂和合成肥料使环境恶化。发现农田中的农药残留和有毒副产物污染了水生生态系统。滥用合成农药不仅影响环境,还影响水生生物的健康状况。有机磷农药污染物是新兴的污染物,威胁着陆地和水生生态系统。久效磷(MCP)是一种有机磷杀虫剂,用于包括水稻在内的作物,玉米,甘蔗,棉花,大豆,花生和蔬菜。MCP本质上是亲水性的,并且它们的增溶性质降低了导致地下水污染的土壤吸附。如果在容器中在38°C下保持稳定,则MCP的半衰期为17-96,并且技术级MCP的半衰期为2500天。MCP引起轻度到重度的混乱,焦虑,过度流涎,哺乳动物和水生动物的惊厥和呼吸窘迫。MCP诱导的毒性包括存活率,行为改变,本综述讨论了不同水生物种的生殖毒性和遗传毒性。此外,这次审查的最终目的是强调国际规则,使用MCP所涉及的未来前景和挑战。
    In recent times, usage of pesticide, herbicides and synthetic fertilizers in farming lands has made the environment worse. The pesticide residues and toxic byproducts from agricultural lands were found to contaminate the aquatic ecosystem. The misuse of synthetic pesticide not only affects the environment, but also affects the health status of aquatic organisms. The organophosphate pesticide pollutants are emerging contaminants, which threatens the terrestrial and aquatic ecosystem. Monocrotophos (MCP) is an organophosphate insecticide, utilized on crops including rice, maize, sugarcane, cotton, soybeans, groundnuts and vegetables. MCP is hydrophilic in nature and their solubilizing properties reduce the soil sorption which leads to groundwater contamination. The half-life period of MCP is 17-96 and the half-life period of technical grade MCP is 2500 days if held stable at 38 °C in a container. MCP causes mild to severe confusion, anxiety, hyper-salivation, convulsion and respiratory distress in mammals as well as aquatic animals. The MCP induced toxicity including survival rate, behavioural changes, reproductive toxicity and genotoxicity in different aquatic species have been discussed in this review. Furthermore, the ultimate aim of this review is to highlight the international regulations, future perspectives and challenges involved in using the MCP.
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  • 文章类型: Case Reports
    Organophosphorus (OP) poisoning is the most common type of poisoning in India. Amongst the OP, monocrotophos poisoning has the highest lethality and need for mechanical ventilation. Monocrotophos is also implicated in causing OP-induced intermediate syndrome, the prevalence of which is 10-40% of all OP poisoning. The other neurological manifestations are delayed neuropathy and neuropsychiatric syndrome. We herein discuss a case of a 58-year-old male who presented with monocrotophos poisoning and intermediate syndrome. During the hospitalisation course, the patient developed hyperammonemic encephalopathy, resulting in difficulty in weaning from mechanical ventilation. After ruling out all possible causes of hyperammonemia, it was attributed to monocrotophos poisoning. The patient improved significantly after initiating lactulose and was successfully weaned off from the ventilator. This report highlights the high index of suspicion of hyperammonemic encephalopathy in monocrotophos toxicity, which can be easily missed due to other commoner neurological manifestations of organophosphorus poisoning.
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  • 文章类型: Journal Article
    表观遗传修饰如DNA甲基化可以改变生物体的表型而不改变其DNA序列。暴露于环境毒物有可能改变水生物种的复原力。然而,关于鱼性腺组织中响应有机磷酸盐的DNA甲基化动力学的信息很少。在目前的工作中,进行减少代表性的亚硫酸氢盐测序以鉴定暴露于有机磷酸盐的Anabastestudienus的卵巢组织中的DNA甲基化模式,特别是久效磷(MCP)。通过测序,平均鉴定出41087个甲基化胞嘧啶位点,并分布在基因的不同部分,即,在转录起始位点(TSS),promotors,外显子,等。在卵巢组织中共检测到1058和1329个差异甲基化区域(DMRs)为高甲基化和低甲基化,分别。利用攀爬鲈鱼的全基因组数据,DMRS,它们相关的重叠基因揭示了外显子内总共22个基因,45个基因在转录起始位点(TSS),和基因间区域的218个基因。通过基因本体分析,共有16个GO术语特别涉及卵泡发育,对氧化应激的反应,卵母细胞成熟,并确定了与生殖生物学相关的多细胞生物对应激的反应。经过功能富集分析,相关的DMG,如类固醇激素生物合成(Cyp19a,11-β-HSD,17-beta-HSD),激素受体(AR,esrrga),类固醇代谢(StAR),孕酮介导的卵母细胞成熟(igf1ar,PGR),与攀援鲈鱼卵巢发育相关的甲基化模式存在显著差异。使用实时PCR对差异甲基化基因(DMG)进行分析,这证明了基因表达水平的改变。这项研究揭示了与化学暴露有关的卵巢发育相关基因的分子水平变化。这项工作为理解DNA甲基化与响应影响水生动物生殖适应性的化学物质的基因调节之间的关系提供了证据。
    Epigenetic modifications like DNA methylation can alter an organism\'s phenotype without changing its DNA sequence. Exposure to environmental toxicants has the potential to change the resilience of aquatic species. However, little information is available on the dynamics of DNA methylation in fish gonadal tissues in response to organophosphates. In the present work, reduced-representation bisulfite sequencing was performed to identify DNA methylation patterns in the ovarian tissues of Anabas testudienus exposed to organophosphates, specifically monocrotophos (MCP). Through sequencing, an average of 41,087 methylated cytosine sites were identified and distributed in different parts of genes, i.e., in transcription start sites (TSS), promoters, exons, etc. A total of 1058 and 1329 differentially methylated regions (DMRs) were detected as hyper-methylated and hypo-methylated in ovarian tissues, respectively. Utilizing whole-genome data of the climbing perch, the DMRs, and their associated overlapping genes revealed a total of 22 genes within exons, 45 genes at transcription start sites (TSS), and 218 genes in intergenic regions. Through gene ontology analysis, a total of 16 GO terms particularly involved in ovarian follicular development, response to oxidative stress, oocyte maturation, and multicellular organismal response to stress associated with reproductive biology were identified. After functional enrichment analysis, relevant DMGs such as steroid hormone biosynthesis (Cyp19a, 11-beta-HSD, 17-beta-HSD), hormone receptors (ar, esrrga), steroid metabolism (StAR), progesterone-mediated oocyte maturation (igf1ar, pgr), associated with ovarian development in climbing perch showed significant differential methylation patterns. The differentially methylated genes (DMGs) were subjected to analysis using real-time PCR, which demonstrated altered gene expression levels. This study revealed a molecular-level alteration in genes associated with ovarian development in response to chemical exposure. This work provides evidence for understanding the relationship between DNA methylation and gene regulation in response to chemicals that affect the reproductive fitness of aquatic animals.
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  • 文章类型: Journal Article
    为了在即时护理(PoC)应用中检测环境和食品样品中的久效磷农药,这项研究,第一次,探索二氧化硅醇凝胶作为固定基质,以支持开发内部定制的纳米功能“chromagrid-lightbox”作为传感系统。该系统使用实验室废料制造,并演示使用智能手机检测高度危险的久效磷农药。纳米发色层是一种芯片状组件,填充有二氧化硅醇凝胶-一种纳米材料(因此称为“纳米发色层”),以及酶促检测久效磷所需的“显色剂”。Lightbox是成像站,可为色栅提供恒定的照明条件,以捕获准确的色度数据。该系统中使用的二氧化硅醇凝胶是通过溶胶-凝胶法由原硅酸四乙酯(TEOS)合成的,并使用先进的分析技术进行了表征。Further,开发了三种色栅测定法,用于光学检测久效磷的低检测限(LOD)为0.421ngml-1(通过α-NAc色栅测定法),0.493ngml-1(通过DTNB铬合金测定)和0.811ngml-1(通过IDA铬合金测定)。开发的新型PoC色栅-灯箱系统能够现场检测环境和食品样品中的久效磷。该系统能够使用可回收的废塑料谨慎地制造。总的来说,这种开发的生态友好型PoC测试系统必将管理环境和可持续农业管理所需的久效磷农药的快速检测。
    With the aim of monocrotophos pesticides detection in environmental and food samples at point-of-care (PoC) application, this research, for the first time, explores silica alcogel as an immobilization matrix to support the development of in-house customized nano-enabled \"chromagrid-lighbox\" as a sensing system. This system is fabricated using laboratory waste materials and demonstrates the detection of highly hazardous monocrotophos pesticide using a smartphone. Nano-enabled chromagrid is a chip-like assembly filled with silica alcogel -a nanomaterial (hence the name \"nano-enabled\" chromagrid), and \"chromogenic reagents\" which is required for the enzymatic detection of monocrotophos. Lightbox is the imaging station fabricated to provide constant lighting conditions to the chromagrid to capture accurate colorimetric data. The silica alcogel used in this system was synthesized from Tetraethyl orthosilicate (TEOS) via a sol-gel method and characterized using advanced analytical techniques. Further, three chromagrid assays were developed for the optical detection of monocrotophos with a low detection limit (LOD) at 0.421 ng ml-1 (by α-NAc chromagrid assay), 0.493 ng ml-1 (by DTNB chromagrid assay) and 0.811 ng ml-1 (by IDA chromagrid assay). The developed novel PoC chromagrid-lightbox system is capable of on-site detection of monocrotophos in environmental as well as food samples. This system is able to be manufacture prudently using recyclable waste plastic. Overall, such developed eco-friendly PoC testing system will surely manage rapid detection of monocrotophos pesticide needed for environmental and sustainable agricultural management.
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  • 文章类型: Journal Article
    Widespread and inadequate use of Monocrotophos has led to several environmental issues. Biodegradation is an ecofriendly method used for detoxification of toxic monocrotophos. In the present study, Msd2 bacterial strain was isolated from the cotton plant growing in contaminated sites of Sahiwal, Pakistan. Msd2 is capable of utilizing the monocrotophos (MCP) organophosphate pesticide as its sole carbon source for growth. Msd2 was identified as Brucella intermedia on the basis of morphology, biochemical characterization and 16S rRNA sequencing. B. intermedia showed tolerance of MCP up to 100 ppm. The presence of opd candidate gene for pesticide degradation, gives credence to B. intermedia as an effective bacterium to degrade MCP. Screening of the B. intermedia strain Msd2 for plant growth promoting activities revealed its ability to produce ammonia, exopolysaccharides, catalase, amylase and ACC-deaminase, and phosphorus, zinc and potassium solubilization. The optimization of the growth parameters (temperatures, shaking rpm, and pH level) of the MCP-degrading isolate was carried out in minimal salt broth supplemented with MCP. The optimal pH, temperature, and rpm for Msd2 growth were observed as pH 6, 35 °C, and 120 rpm, respectively. Based on optimization results, batch degradation experiment was performed. Biodegradation of MCP by B. intermedia was monitored using HPLC and recorded 78% degradation of MCP at 100 ppm concentration within 7 days of incubation. Degradation of MCP by Msd2 followed the first order reaction kinetics. Plant growth promoting and multi-stress tolerance ability of Msd2 was confirmed by molecular analysis. It is concluded that Brucella intermedia strain Msd2 could be beneficial as potential biological agent for an effective bioremediation for polluted environments.
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  • 文章类型: Journal Article
    已知的有机磷农药广泛用于农业中以改善作物的生产。根据文献,对某些有机磷农药的降解进行了理论研究。然而,在环境中,甲硫磷和久效磷降解过程中的毒性机制和变化尚不清楚,尤其是在废水中。在这项研究中,两种代表性有机磷农药降解的反应机理(即,使用量子化学方法wB97-XD/6-311++G(3df,2pd)//wB97-XD/6-311++G(d,P).结果表明,主要通道是在CC键中C原子上的OH加成,能垒分别为15.6和14.7kJ/mol,在大气和水中,分别,给mepinphos.至于久效磷,在大气和水中,通过8.2和10.6kJ/mol的屏障从NH基团提取H更可行。此外,还研究了大气中主要产物与NO和O2的后续反应,以评估mepinphos和久效磷的大气化学。动力学上,大气中的速效磷和久效磷的总速率常数为2.68×10-9和3.86×10-8cm3-1·s-1,而在298K的水中为4.91×1010和7.77×1011M-1s-1,因此,在大气中的寿命估计为36.46-364.60s(2.53-25.31s),在高级氧化过程(AOPs)系统中,和1.41×10-2-1.41×10-1s(8.92×10-4-8.92×10-3s)。此外,对大鼠和三种水生生物的生态毒性预测表明,通过使用ECOSAR和基于T.E.S.T程序的定量结构和活性关系(QSAR)方法,可以降低其降解过程中的毒性。
    Known organophosphorus pesticides are used widely in agriculture to improve the production of crops. Based on the literature, the degradation of some organophosphorus pesticides was studied theoretically. However, the mechanisms and variation of toxicity during the degradation of mevinphos and monocrotophos are still unclear in the environment, especially in wastewater. In this study, the reaction mechanisms for the degradation of the two representative organophosphorus pesticides (i.e., mevinphos and monocrotophos) in presence of OH radicals in the atmosphere and water are proposed using quantum chemical methods wB97-XD/6-311 + +G(3df,2pd)//wB97-XD/6-311 + +G(d,p). Result shows that the dominant channel is OH-addition to the C atom in CC bond with energy barriers being 15.6 and 14.7 kJ/mol, in the atmosphere and water, respectively, for mevinphos. As for monocrotophos, H-abstraction from NH group via barriers of 8.2 and 10.6 kJ/mol is more feasible in both the atmosphere and water. Moreover, the subsequent reactions of the major products in the atmosphere with NO and O2 were also studied to evaluate the atmospheric chemistry of mevinphos and monocrotophos. Kinetically, the total rate constant is 2.68 × 10-9 and 3.86 × 10-8 cm3 molecule-1·s-1 for mevinphos and monocrotophos in the atmosphere and 4.91 × 1010 and 7.77 × 1011 M-1 s-1 in the water at 298 K, thus the lifetime is estimated to be 36.46-364.60 s (2.53-25.31 s) in the atmosphere, and 1.41 × 10-2 - 1.41 × 10-1 s (8.92 ×10-4 - 8.92 ×10-3 s) in the advanced oxidation processes (AOPs) system. Furthermore, ecotoxic predictions for rats and three aqueous organisms imply their toxicity are reduced during degradation by using ECOSAR and T.E.S.T program based quantitative structure and activity relationship (QSAR) method.
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  • 文章类型: Journal Article
    钾(K)和磷(P)是植物发育所需的重要营养元素,但它是广泛存在的不溶性形式为植物的吸收。为了提高生产力,生物肥料在促进植物生长中起着至关重要的作用。我们目前的工作重点是从番茄栽培土壤的农业土壤中分离磷酸钾溶解细菌。用分光光度法估算久效磷的钾和磷酸盐溶解和降解。在13个分离株中,两个分离株被证明是最好的P和K增溶剂。对细菌分离株(SDKVG02和SDKVG04)进行了优化,以获得分离株对57.5mgL-1和15.07mgL-1的最大P和K溶解。盆栽实验使用固定在载体材料上的SDKVG02和04进行,泥炭是最佳载体,总平均绿克和鹰嘴豆长度为11.66±0.066622.22±0.0577。通过SDKVG02和SDKVG04,在80ppm的MCP下实现MCP降解百分比,具有75.8%和64.10%。此外,生产有机酸,如苹果酸,邻苯二甲酸,抗坏血酸,烟酸,根据16SrRNA基因测序,将分离株鉴定为血球肠杆菌-SDKVG-02,阴沟肠杆菌SDKVG-04。KSB-PSB分离株还表达N-固定活性,这通过计算机分析得到证明。值得强调的是SDKVG02和04将是有效的生物肥料,可用于提高土壤肥力和作物生产力以及土壤中久效磷的降解。
    Potassium (K) and phosphorus (P) are the important macronutrients needed for the plant development, but it is widely present in an insoluble form for the plant\'s uptake. In order to increase the productivity, biofertilisers play crucial role in plant growth enhancement. Our present work focused to isolate potassium-phosphate solubilizing bacteria from the agricultural soil of tomato cultivated soil. Potassium and phosphate solubilization and degradation of monocrotophos was estimated spectrophotometrically. Out of thirteen isolates, two isolates proved to be the best P and K solubilizers. The bacterial isolates (SDKVG02 and SDKVG04) were optimized to obtain maximum P and K solubilization of 57.5 mg L-1 and 15.07 mg L-1 by the isolates. Pot experiments were conducted using SDKVG 02 and 04, immobilized on carrier materials, peat proving the best carrier with the total average green gram and chick pea length of 11.66 ± 0.0666 22.22 ± 0.0577. The MCP degradation percentage was achieved at 80 ppm of MCP with 75.8% and 64.10% by SDKVG 02 and SDKVG 04. Furthermore, production of organic acids such as malic acid, phthalic acid, ascorbic acid, nicotinic acid, and tartaric acid paves solubilization of P and K. The isolates were recognized based on 16S rRNA gene sequencing as Enterobacter hormaechei- SDKVG-02, Enterobacter cloacae SDKVG- 04. The KSB-PSB isolates also express N-fixing activity which is proved through In-silico analysis. It is worth to highlight SDKVG 02 and 04 would be potent biofertiliser exploited in increasing the soil fertility and crop productivity as well in degradation of monocrotophos present in the soil.
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  • 文章类型: Journal Article
    暴露于有机磷农药(OP)可能会产生慢性不良反应,这些不良反应与乙酰胆碱酯酶的抑制作用无关。急性OP毒性的经典目标。在纯蛋白质中,有机磷农药毒死rifoxon诱导赖氨酸和谷氨酸(或天冬氨酸)之间的交联,并失去水分。微管蛋白对OP诱导的交联特别敏感。我们的目标是探索复杂蛋白质样品中OP诱导的交联,来自Susscrofa的富含MAP的微管蛋白,并测试8OP促进异肽交联的能力。我们用100μM毒死rif处理了100μgMAP丰富的微管蛋白,毒死蜱oxon,甲胺磷,对氧磷,二嗪农,重氮,久效磷,或者敌敌畏.使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离每个样品并用考马斯蓝染色。五个凝胶切片(约30、50、150和300kDa,和分离凝胶的顶部)从八个OP样品中的每一个的泳道和未处理的对照泳道中移出。对这些凝胶切片进行凝胶内胰蛋白酶消化。检查胰蛋白酶肽的MSMS片段化谱的异肽交联。16个光谱为异肽交联肽提供了令人信服的证据。十种来自毒死蜱的oxon反应,1来自敌敌畏,1来自对氧磷,1来自二嗪农,和3来自二氮唑。结论是,蛋白质交联的催化作用是有机磷农药和农药代谢产物的一般性质。数据可通过ProteomeXchange获得,标识符为PXD034529。
    Exposure to organophosphorus pesticides (OP) can have chronic adverse effects that are independent of inhibition of acetylcholinesterase, the classic target for acute OP toxicity. In pure proteins, the organophosphorus pesticide chlorpyrifos oxon induces a cross-link between lysine and glutamate (or aspartate) with loss of water. Tubulin is particularly sensitive to OP-induced cross-linking. Our goal was to explore OP-induced cross-linking in a complex protein sample, MAP-rich tubulin from Sus scrofa and to test 8 OP for their capacity to promote isopeptide cross-linking. We treated 100 μg of MAP-rich tubulin with 100 μM chlorpyrifos, chlorpyrifos oxon, methamidophos, paraoxon, diazinon, diazoxon, monocrotophos, or dichlorvos. Each sample was separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie blue. Five gel slices (at about 30, 50, 150, and 300 kDa, and the top of the separating gel) were removed from the lanes for each of the eight OP samples and from untreated control lanes. These gel slices were subjected to in-gel trypsin digestion. MSMS fragmentation spectra of the tryptic peptides were examined for isopeptide cross-links. Sixteen spectra yielded convincing evidence for isopeptide cross-linked peptides. Ten were from the chlorpyrifos oxon reaction, 1 from dichlorvos, 1 from paraoxon, 1 from diazinon, and 3 from diazoxon. It was concluded that catalysis of protein cross-linking is a general property of organophosphorus pesticides and pesticide metabolites. Data are available via ProteomeXchange with identifier PXD034529.
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