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嗜根寡养单胞菌(Stenotrophomonas rhizophila)JC1重金属抗性及其解毒蛋白生物信息学分析 被引量:2
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作者 孙尚琛 陈吉祥 +2 位作者 王永刚 陈凯 冷非凡 《安全与环境学报》 CAS CSCD 北大核心 2021年第4期1788-1796,共9页
为研究某些重金属解毒蛋白在微生物抗重金属过程中发挥的作用,从常年受重金属污染的某化工厂附近土壤中分离、驯化得到一株对铅、铜、镉3种金属有较好抗性的嗜根寡养单胞菌(Stenotrophomonas rhizophila)JC1,并对其进行了全基因组测序... 为研究某些重金属解毒蛋白在微生物抗重金属过程中发挥的作用,从常年受重金属污染的某化工厂附近土壤中分离、驯化得到一株对铅、铜、镉3种金属有较好抗性的嗜根寡养单胞菌(Stenotrophomonas rhizophila)JC1,并对其进行了全基因组测序。通过分析其蛋白家族、代谢通路等,发现蛋白质AHY58868.1、AHY59763.1、AHY58405.1和AHY57635.1与重金属解毒直接相关,故利用分析软件对4种蛋白的生物学性质、结构功能和相互作用等分别进行了分析。生理生化试验结果表明,菌株JC1表面光滑,菌落边缘整齐,为无芽孢的革兰氏阳性菌,Pb^(2+)、Cu^(2+)和Cd^(2+)的去除率最高分别可达76.9%、96.7%和47.7%。生物信息学分析结果表明,AHY58405.1为不稳定的亲水性蛋白,而其余3种为相对较稳定的亲水性蛋白。AHY58868.1和AHY59763.1无信号肽结构,而AHY58405.1和AHY57635.1存在信号肽结构。AHY58868.1和AHY57635.1于细胞内膜发挥作用,AHY59763.1在细胞质内发挥作用,AHY58405.1在周质空间发挥作用。蛋白质结构功能和蛋白间相互作用关系分析表明,AHY58868.1主要参与机体铜离子的转运,对维持细胞内铜平衡起关键作用;AHY59763.1和AHY58405.1主要介导重金属与某些阴离子(—PO4^(3-)、—SH、—NH_(2)、—COOH等)基团的络合,尤其是AHY59763.1在微生物体内可为二价重金属的络合提供多个活性位点;AHY57635.1主要参与机体的ABC转运蛋白的转运过程,可介导重金属向胞外的排出。 展开更多
关键词 环境科学技术基础学科 嗜根寡养单胞菌 解毒蛋白 生物信息学 结构 功能
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眼镜蛇血清解毒蛋白基因的分子克隆和序列分析 被引量:2
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作者 王晓路 邵靖宇 《中国医学科学院学报》 CAS CSCD 北大核心 1996年第4期263-272,共10页
既往研究中作者发现,眼镜蛇血清解毒蛋白(CSAP)经胰蛋白酶水解后纯化所得的几个肽段中,至少在一39肽显示其序列与大鼠血清白蛋白相关。为进一步弄清CSAP的毒素结合活性与其相应的分子结构关系,以眼镜蛇肝细胞建立了cD... 既往研究中作者发现,眼镜蛇血清解毒蛋白(CSAP)经胰蛋白酶水解后纯化所得的几个肽段中,至少在一39肽显示其序列与大鼠血清白蛋白相关。为进一步弄清CSAP的毒素结合活性与其相应的分子结构关系,以眼镜蛇肝细胞建立了cDNA文库,借已知的胰蛋白酶水解CSAP所得的肽段氨基酸序列设计了一组PCR引物。采用PCR扩增方法从文库中筛选得CSAP特异性克隆,测定了CSAP全cDNA序列,然后推导出CSAP氨基酸序列。并得知CSAP为一由614个氨基酸残基组成的单一多肽链,分子量69800。根据其全序列的结构特征,尤其是其信号肽与已知的几种哺乳动物白蛋白的信号肽序列非常一致,其中半胱氨酸残基的分布规律也与其他血清白蛋白非常相似,可确认CSAP即为眼镜蛇血清白蛋白(CSA)。 展开更多
关键词 眼镜蛇 血清 解毒蛋白 CDNA 文库 克隆 蛋白
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眼镜蛇血清解毒蛋白的溴化氰裂解肽段与自身神经毒素的结合
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作者 柴婴蕾 邵靖宇 《中国医学科学院学报》 CAS CSCD 北大核心 1999年第5期395-398,共4页
目的为进一步认识眼镜蛇血清解毒蛋白(CSAP)如何与神经毒素结合并解毒。方法用溴化氰裂解CSAP肽链中由Met羧基端组成的肽键,裂解产物经超滤除去分子量小于10000的片段后,用亲和层析法得到与毒素仍保持有亲和力的肽... 目的为进一步认识眼镜蛇血清解毒蛋白(CSAP)如何与神经毒素结合并解毒。方法用溴化氰裂解CSAP肽链中由Met羧基端组成的肽键,裂解产物经超滤除去分子量小于10000的片段后,用亲和层析法得到与毒素仍保持有亲和力的肽段。结果与毒素仍保持有亲和力的肽段相当于Lys2-Met485,Lys2-Met275,Lys276-Met485,Pro444-Met603。结论CSAP的3个结构域均具有与毒素分子结合的活性,CSAP结合的8个毒素分子分散在整个肽链的各个段落上。 展开更多
关键词 眼镜蛇 解毒蛋白 CSAP 神经毒素 溴化氰 裂解
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绿原酸对美国白蛾幼虫生长发育和解毒相关蛋白活性的影响 被引量:7
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作者 潘忠玉 莫夏娜 +1 位作者 孟香 陈敏 《昆虫学报》 CAS CSCD 北大核心 2020年第9期1081-1090,共10页
【目的】明确植物次生代谢物质绿原酸对美国白蛾Hyphantria cunea幼虫生长发育和解毒相关蛋白活性的影响。【方法】将含不同浓度绿原酸(0,0.125%,0.250%,0.500%,1.000%和2.000%)的人工饲料饲养美国白蛾幼虫,测定各组中5龄幼虫6 d内的死... 【目的】明确植物次生代谢物质绿原酸对美国白蛾Hyphantria cunea幼虫生长发育和解毒相关蛋白活性的影响。【方法】将含不同浓度绿原酸(0,0.125%,0.250%,0.500%,1.000%和2.000%)的人工饲料饲养美国白蛾幼虫,测定各组中5龄幼虫6 d内的死亡率,各组中5龄幼虫在取食48 h时的营养效应指标,0.500%绿原酸处理组中幼虫(3龄幼虫开始)的生长发育,以及在取食36 h时各组中5龄幼虫肠道中的解毒相关蛋白活性。【结果】绿原酸处理组中美国白蛾5龄幼虫6 d内的死亡率随着人工饲料中绿原酸浓度的增大而逐渐升高,且显著高于对照组(取食含10%DMSO人工饲料的)。绿原酸对美国白蛾5龄幼虫的营养效应指标有显著影响,各处理组幼虫的相对取食量和近似消化率显著高于对照组,食物转化率显著低于对照组,除0.125%绿原酸处理组外,其余处理组幼虫的食物利用率和相对生长速率也显著低于对照组。0.500%绿原酸处理使美国白蛾3-5龄幼虫发育历期比对照组显著延长28.24%(3.7 d),老熟幼虫(6-7龄幼虫)的发育历期缩短8.97%(0.7 d),且幼虫总成活率、化蛹率、羽化率、雌雄性比及产卵量等各项指标均显著低于对照组。不同浓度绿原酸处理对美国白蛾幼虫肠道中的细胞色素P450酶(CYP450)、谷胱甘肽S-转移酶(GSTs)、尿苷二磷酸葡萄糖醛酸转移酶(UGT)和ABC转运蛋白(ABC transporters)的活性具有显著影响,但对羧酸酯酶(CarE)活性未见显著影响。【结论】绿原酸可影响美国白蛾幼虫的存活率、食物利用效率、生长发育和繁殖,而美国白蛾幼虫可能通过调节食物利用率和诱导其解毒相关蛋白活性从而对食物中绿原酸产生适应性。 展开更多
关键词 美国白蛾 绿原酸 营养效应 生长发育 解毒相关蛋白
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黄连解毒汤对冠心病心绞痛患者C反应蛋白和肿瘤坏死因子-α的影响 被引量:15
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作者 朱娜 曹雪明 +2 位作者 蔡颖娴 吴玉 汪彬彬 《中国中医急症》 2012年第4期542-543,共2页
目的观察黄连解毒汤对冠心病心绞痛患者C反应蛋白(CRP)和肿瘤坏死因子-α(TNF-α)的影响。方法选取已确诊冠心病患者110例,随机分为治疗组和对照组,对照组采用西医常规治疗;治疗组加用经典方剂黄连解毒汤。结果 治疗后两组CRP和TNF-α... 目的观察黄连解毒汤对冠心病心绞痛患者C反应蛋白(CRP)和肿瘤坏死因子-α(TNF-α)的影响。方法选取已确诊冠心病患者110例,随机分为治疗组和对照组,对照组采用西医常规治疗;治疗组加用经典方剂黄连解毒汤。结果 治疗后两组CRP和TNF-α水平均下降,治疗组优于对照组。结论黄连解毒汤可以通过降低CRP与TNF-α水平抑制炎症反应以改善内皮细胞功能,进而减轻冠状动脉粥样硬化,改善心肌缺血缺氧,缓解冠心病心绞痛患者症状。 展开更多
关键词 冠心病心绞痛 黄连解毒汤C反应蛋白 肿瘤坏死因子-Α
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星天牛转录组及三大解毒酶家族相关基因系统发育分析 被引量:1
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作者 韩小红 卢赐鼎 +5 位作者 华银 林浩宇 是雨霏 吴松青 张飞萍 梁光红 《林业科学》 EI CAS CSCD 北大核心 2019年第5期104-113,共10页
【目的】研究星天牛转录组信息及体内细胞色素P450单加氧酶(cytochrome P450 monooxygenase,CYP)、羧酸酯酶(carboxylesterase,CarE)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)三大解毒酶系的表达谱情况,通过构建系统进化树,... 【目的】研究星天牛转录组信息及体内细胞色素P450单加氧酶(cytochrome P450 monooxygenase,CYP)、羧酸酯酶(carboxylesterase,CarE)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)三大解毒酶系的表达谱情况,通过构建系统进化树,探析其这3种酶系基因家族的类别及其系统演化关系,为星天牛对多寄主种类的适应性与杀虫剂的抗药性机制研究提供依据。【方法】采用新一代高通量测序技术Illumina HiSeq TM 2500 对星天牛进行转录组测序与数据分析,通过数据筛选并进一步与鞘翅目近缘物种的解毒酶同源蛋白进行系统发育分析。【结果】经测序、序列拼接得到55 260条unigenes,将其与公共数据库进行同源比对,注释了32 247条unigenes,在Swiss-Prot数据库注释的unigenes的数量最多(99.11%);注释到Nr数据库时与赤拟谷盗的unigenes同源性最高(60.25%);在GO数据库中,共有8 083个unigenes被成功注释,根据其功能大致可分为3 类47 亚类;在KEGG数据库中,4 600 条unigenes与162个预测路径可以匹配。通过基因注释发现248条解毒酶系基因在星天牛体内表达,包括139条CYP基因、72条CarE基因、37条GST基因。系统发育分析发现,星天牛的大部分CYP基因都至少与一种鞘翅目近缘物种的CYP基因聚类在一起,P450基因中的CYP6家族数量与其他近缘物种(鞘翅目)类似,包含的基因数量最多;CarE解毒酶蛋白基因主要为β-酯酶、神经趋化蛋白、外源性代谢酶、未知等4种类型,分别可能参与星天牛的一些激素以及信息素的加工、神经和发育过程、消化与解毒等生理过程;星天牛的GST基因聚类主要包含Microsomal GST、Sigma GST、Omega GST、Delta GST、Theta GST等亚家族,具有保护生物大分子免受氧化损伤、催化活力等功能。【结论】本研究获得星天牛的转录组信息,初步探明星天牛体内三大解毒酶系的表达谱及分化情况,可为星天牛对多寄主种类的适应性与杀虫剂抗药性的产生及遗传机制研究提供基础数据和参考。 展开更多
关键词 星天牛 转录组 高通量测序 系统发育 解毒蛋白基因
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Research Progress on Heavy Metals Detoxification in Human Body 被引量:2
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作者 张辉 孙建霞 +5 位作者 李名薇 朱翠娟 胡云峰 吴实 焦睿 白卫滨 《Agricultural Science & Technology》 CAS 2015年第8期1785-1790,1809,共7页
With the pollution of heavy metals becoming more and more serious, hu- man health suffers from great harms, making the detoxification in human body admit of no delay. This paper summarized the heavy metals used for de... With the pollution of heavy metals becoming more and more serious, hu- man health suffers from great harms, making the detoxification in human body admit of no delay. This paper summarized the heavy metals used for detoxification in hu- man body in recent years, including metallothionein, polyphenols, dietary fibers from seaweed, GTS, and explored the detoxification mechanism of these heavy metals in human body. 展开更多
关键词 Heavy metal Detoxification in human body Metaltothionein ANTHOCYANIN
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Identification of the immunogenic domains in HBsAg preS1 region using overlapping preS1 fragment fusion proteins 被引量:7
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作者 Wei-GuoHu JunWei Heng-ChuanXia Xin-XiuYang FengLi Guang-DiLi YuanWang Zu-ChuanZhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第14期2088-2094,共7页
AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purp... AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitopebased hepatitis B vaccine design.METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.RESULTS: The results in mice showed that the immunogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity.More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic domains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitopebased HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans. 展开更多
关键词 HBV preSl GST fusion protein Immunogenic domain OVERLAPPING
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Functional interplay among the flavivirus NS3 protease, helicase, and cofactors 被引量:2
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作者 Kuohan Li Wint Wint Phoo Dahai Luo 《Virologica Sinica》 CAS CSCD 2014年第2期74-85,共12页
Flaviviruses are positive-sense RNA viruses, and many are important human pathogens. Nonstructural protein 2B and 3 of the flaviviruses(NS2BNS3) form an endoplasmic reticulum(ER) membrane-associated hetero-dimeric com... Flaviviruses are positive-sense RNA viruses, and many are important human pathogens. Nonstructural protein 2B and 3 of the flaviviruses(NS2BNS3) form an endoplasmic reticulum(ER) membrane-associated hetero-dimeric complex through the NS2B transmembrane region. The NS2BNS3 complex is multifunctional. The N-terminal region of NS3, and its cofactor NS2B fold into a protease that is responsible for viral polyprotein processing, and the C-terminal domain of NS3 possesses NTPase/RNA helicase activities and is involved in viral RNA replication and virus particle formation. In addition, NS2BNS3 complex has also been shown to modulate viral pathogenesis and the host immune response. Because of the essential functions that the NS2BNS3 complex plays in the flavivirus life cycle, it is an attractive target for antiviral development. This review focuses on the recent biochemical and structural advances of NS2BNS3 and provides a brief update on the current status of drug development targeting this viral protein complex. 展开更多
关键词 crystal structures antiviral drug target serine protease RNA helicase
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Molecular Dissection of Bombyx mori Nucleopolyhedrovirus orf8 Gene
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作者 WonKyung Kang 《Virologica Sinica》 SCIE CAS CSCD 2009年第4期315-322,共8页
Viruses including baculoviruses are obligatory parasites, as their genomes do not encode all the proteins required for replication. Therefore, viruses have evolved to exploit the behavior and the physiology of their h... Viruses including baculoviruses are obligatory parasites, as their genomes do not encode all the proteins required for replication. Therefore, viruses have evolved to exploit the behavior and the physiology of their hosts and olden coevolved with their hosts over millions of years. Recent comparative analyses of complete genome sequences of baculoviruses revealed the patterns of gene acquisitions and losses that have occurred during baculovirus evolution. In addition, knowledge of virus genes has also provided understanding of the mechanism of baculovirus infection including replication, species-specific virulence and host range. The Bm8 gene of Bombyx mori nucleopolyhedrovirus (NPV) and its homologues are found only in group I NPV genomes. The Autographa californica NPV Ac 16 gene is a homologue of Bm8 and, encodes a viral structural protein. It has been shown that BmS/Ac 16 interacts with baculoviral and cellular proteins. BmS/Ac 16 interacts with baculoviral IE1 that is facilitated by coiled coil domains, and the interaction with IE1 is important for Bin8 function. Acl6 also forms a complex with viral FP25 and cellular actin and associates with membranes via palmitoylation. These data suggested that this gene family encodes a multifunctional protein that accomplishes specific needs of group I NPVs. 展开更多
关键词 Group I NPV Bm8 Early gene Structural protein Nuclear localization
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Purification and characterization of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China
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作者 余晓东 李博 余争平 《Journal of Medical Colleges of PLA(China)》 CAS 2005年第3期129-134,共6页
Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated fr... Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated from the venom of Chinese Trimeresurus mucrosqumatus by a combination of ion-exchange chromatography on DEAE-Sephadex A-50, CM-Sepharose Cl-6B and gel filtration on Sephadex G-100.Results: The purified protein named TM-AEH,a glycoprotein with carbohydrate content of 0.5% neutral hexose and 0.75% sialic acid,a relative molecular mass of 29.0 kDa,and an isoelectric point(pI) of 5.2. It shares with an extinction coefficient(E 0.1%/cm) of 1.332 at 280 nm,consisted of 225 amino acid residues,and migrated as a band under reduced or non-reduced condition in basic PAGE.TM-AEH was a highly thermostable protein and was stable to pH changes between 5 and 9.The optimum temperature and optimum pH were 55℃ and 8.4 for its catalytic activity respectively,which was inhibited by Fe 3+ and Cu 2+.Conclusion:This protein can exhibit higher BAEE-hydrolysing activity and fibrinogenolytic activity as compared to that of whole venom. 展开更多
关键词 Trimeresurus mucrosqumatus snake venom arginine ester-hydrolysing enzyme fibrinogenolytic activities
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Detoxification of toxic herbs in TCM prescription based on modulation of efflux transporters
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作者 QIAN Liyunhe HE Yufei +1 位作者 ZHANG Mei XIE Ying 《Digital Chinese Medicine》 2021年第1期9-18,共10页
Traditional Chinese medicines(TCMs)have been used to prevent and treat various diseases for thousands of years.Promoting efficacy and reducing toxicity by the compatibility theory of TCMs has attracted increasing atte... Traditional Chinese medicines(TCMs)have been used to prevent and treat various diseases for thousands of years.Promoting efficacy and reducing toxicity by the compatibility theory of TCMs has attracted increasing attention,especially for the toxicity of herbs.Studies have pointed out the interactions between the active compounds of herbs and transporters in the detoxification process of toxic compounds.Here,we summarize data on five toxic herbs commonly used in TCMs and their related efflux transporters to reduce toxicity to offer a scientific rationale for the compatibility principle of TCMs and provide guidance for the rational clinical use of TCMs. 展开更多
关键词 Toxic herbs Efflux transporters DETOXIFICATION TCMs compatibility principles P-glycoprotein(P-gp)
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