Structure and mechanism of copper, zinc superoxide dismutase

Copper, zinc superoxide dismutase (SOD) catalyses the very rapid two-step dismutation of the toxic superoxide radical (O-2) to molecular oxygen and hydrogen peroxide through the alternate reduction and oxidation of the active-site copper. We report here that after refitting and further refinement of the previous 2 A structure of SOD2, analysis of the new model and its calculated molecular surface shows an extensive surface topography of sequence-conserved residues stabilized by underlying tight packing and H-bonding. There is a single, highly complementary position for O-2 to bind to both the Cu(II) and activity-important Arg 141 with correct geometry; two water molecules form a ghost of the superoxide in this position. The geometry and molecular surface of the active site, together with biochemical data, suggest a specific model for the enzyme mechanism.     

SDS、Cu~(2+)复合污染对蟾蜍蝌蚪血红细胞核异常及抗氧化酶活性的影响

以中华大蟾蜍蝌蚪为试验对象,采用暴露试验法,研究了亚急性条件下十二烷基硫酸钠(SDS)、Cu2+单一及复合污染对蝌蚪血红细胞核异常率及肝脏超氧化物歧化酶(SOD)活性的影响。结果表明:单一SDS、Cu2+处理条件下,随着SDS、Cu2+暴露浓度增加,蝌蚪血红细胞核异常率及肝脏的抗氧化酶SOD与对照组相比均呈浓度-效应关系;SDS、Cu2+复合处理条件下,污染物对蝌蚪肝脏的SOD活性及血红细胞核异常率的影响表现为一定的协同作用,但污染物不同质量浓度的组合对SOD活性及血红细胞核异常率影响程度不同。     

银杏种子脱水敏感性的研究

【目的】研究银杏种子脱水敏感性及相关的生理生化变化,为银杏种子贮藏、种质资源的保存及苗木生产提供理论依据。【方法】银杏种子于25 ℃、湿度(25±2)%条件下脱水67 d,种子含水量每下降5%左右时随机取样测定相关生理生化指标。【结果】当种子含水量降至45.1%时,发芽率并未有明显变化,然而,当种子含水量降至40.1%时,发芽率从开始的92%下降至50%;相关性分析表明种子含水量与发芽率之间存在极显著的正相关(r=0.910)。银杏种子脱水过程中,其相对电导率不断上升。此外,SOD活性变化与POD活性的变化基本相似,均呈现出先下降,后升高至最大值,再下降的趋势,只是其POD活性的上升比SOD早,说明POD的反应比SOD更为敏感。种子含水量与硫代巴比妥酸反应产物(TBARS)含量(r=-0.619)、相对电导率(r=-0.745)之间存在极显著的负相关关系。【结论】银杏种子对脱水高度敏感,表现出顽拗型种子特点。严重脱水降低了银杏种子抗氧化酶活性,并加剧了膜脂过氧化作用,导致种子活力下降乃至完全丧失。     

转人Cu,Zn-SOD基因马铃薯的研究

采用RT—PCR技术从人肝总RNA中分离扩增了490bp的人铜锌超氧化物歧化酶(hCM，Zn—SOD)基因的cDNA序列，进行了序列测定，结果和文献报道的一致；构建了CaMv35S启动子驱动hCu，Zn—SOD基因的植物表达载体PBICuSOD；用三亲交配法将重组质粒PBICuSOD转化到农杆菌LBA4404，转化马铃薯得到转基因植株。PCR和Southern—blot分析证明，hCu，Zn—SOD基因巳整合到马铃薯基因组中。Western blot分析表明，hCu，Zn—SOD基因在转基因马铃薯植株中得到表达。用NBT光还原法测定转基因马铃薯植株中的SOD酶活力，叶的总酶活为1066．6U／g(湿重，下同)，块茎的总酶活为10llU／g。     

硅对盐胁迫下金丝小枣3种抗氧化酶的影响

以1年生金丝小枣嫁接苗为试材,研究硅对盆栽苗在盐胁迫下谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性的影响.结果表明:在盐浓度(NaCl质量分数)小于0.4%时,SOD和GSH-Px的活性随着土壤含盐量的增加而增加,POD活性在NaCl质量分数为0.25%的盐胁迫下升高,在NaCl质量分数为0.4%的盐胁迫时下降;盐胁迫下加硅可以显著提高金丝小枣SOD、POD和GSH-Px的活性且差异显著.笔者认为盐胁迫下给土壤施硅可以缓解盐分对金丝小枣的毒害作用.     

甜瓜叶片的衰老与超氧化物歧化酶活性及脂质过氧化作用的关系

分别用全营养液、缺Cu-Zn营养液、缺Fe营养液、缺Mn营养液、10％NaCl营养液培养甜瓜幼苗,研究不同时间甜瓜叶片的超氧化物歧化酶(SOD)及脂质过氧化产物丙二醛(MDA)含量的变化。实验结果表明：随着胁迫时间的延长,SOD活性降低,MDA含量显著增高。证明甜瓜为Cu-Zn SOD。观察了SOD同工酶随胁迫时间的延长在叶片中的变化。讨论了离子胁迫过程中氧自由基对酶及质膜的损伤影响。     

Superoxide involvement in the bactericidal effects of negative air ions on Staphylococcus albus.

The physical nature of small air ions is well established and it is recognized that they can produce a variety of biological effects. However, in only a few instances have any underlying biochemical changes been detected. Theoretically, one can consider the hydrated superoxide radical anion (O2) (H2O)n with n congruent to 4-8 as a likely candidate for a biologically active species of negative air ion. The chemical and biological reactivity of superoxide is high and includes a leading role in bacterial killing caused by radiation, in which superoxide dismutase (SOD), an enzyme that catalyses the reaction: O2 + O2 +2H leads to H2O2 +O2 protected markedly. Other studies have also demonstrated the bactericidal effect of O2 (refs 9-11). Inasmuch as the bactericidal action of small negative air ions has been repeatedly confirmed, we decided to test for the involvement of O2 in this phenomenon by evaluating the protective effect of SOD. Our results show strong O2 involvement in negative air ion bacterial kill.     

离子交换色谱柱上橙皮苷分离与配位反应的一体化

提出并实现了橙皮苷分离及其与Cu2 配位反应一体化的新设想,并借助离子交换色谱、紫外可见光谱、X-射线衍射、傅立叶红外光谱和核磁共振等分析手段探讨其作用机理.结果表明,在离子交换色谱柱上能有效实现配体分离与配位反应的一体化,制备高生物活性中药配合物新药.其反应机理在于树脂相中软碱橙皮苷与溶液相中交界酸Cu2 间存在库仑力,容易发生配位反应,配体的分子状态和疏水状态因Cu2 对橙皮素的"集中效应"和芸香二糖对橙皮素的"包埋效应"而改变,故再生剂阴离子易穿过能斯特层与橙皮苷交换,并因电子云重叠而排斥橙皮苷至液相中,从而有助于分离、反应一体化.     

瑞香素及其铜(Ⅱ)、锌(Ⅱ)配合物对超氧自由基的清除作用

本文以健康人血红细胞及细胞膜为实验材料,利用核黄素-蛋氨酸光照体系产生O_D~-,考察了瑞香素及其铜(Ⅱ)、锌(Ⅱ)配合物时O_2~-的清除作用。结果表明:瑞香素及其铜(Ⅱ)、锌(Ⅱ)配合物对O_2~-都有明显清除作用,以钢(Ⅱ)配合物活性最高;O_2~-可使Hb氧化,使膜中过氧化脂质含量增加,SOD、瑞香素及其铜(Ⅱ)、锌(Ⅱ)配合物对此均有一定的抑制作用,其中以SOD活性最高。     

超氧化物歧化酶的循环伏安行为

采用循环伏安方法研究了超氧化物歧化酶在不同材料电极上的电化学行为，重点报道了在金微盘电极上的电子迁移过程，并计算了电化学反应的参数．     

静电场对苜蓿幼苗期酶活性的影响

为了研究静电对紫花苜蓿(Medicago sativa Leguminosae)抗旱性的影响，寻找出提高紫花苜蓿幼苗抗旱性的最佳电场条件。在静电场不同电场强度下处置紫花苜蓿种子10min后，室温下培养9d，再测定幼苗的丙二醛(MDA)的含量和过氧化物酶(POD)、超氧化物酶(SOD)的活性。实验所用电极电压波形为50Hz半波整流，电场强度共11个，其中1个未经电场处理作为对照组。结果表明，在11个电场处理的材料中，有6个MDA水平低于对照，1个与对照相同，其余4个高于对照，变化幅度为22．8％～34．8％。SOD酶和POD酶的活性普遍降低，降低幅度分别为7．7％～57．7％和0．5％～59．4％。静电场处理可以提高紫花苜蓿幼苗的抗旱性。     

一类非线性反应扩散系统正解的存在性

利用上下解原理的方法，研究一类生态学中的反应扩散系统Volterra-Lotka模型的共生态。讨论了等出生率情形的互助模型正解的分歧结构，给出了正解的存在性，不存在性和惟一性条件。结果表明：对这类反应扩散方程组的理解依赖于对单个半线性椭圆方程的理解。     

自制纤维素金属螯合亲和膜的部分吸附性能及纯化牛血SOD

×10-6 g/cm2,但蛋白解吸率前者低于后者,表明一定浓度的NaCl有助于亲和膜对蛋白质的吸附,同时也使被吸附的蛋白的解吸率有所降低.吸附-解吸方式(静态法)分离纯化SOD,解吸样品比活达8 770 U/mg,纯度为81%;吸附-洗脱方式(动态法)分离纯化SOD,穿柱流出样品比活达12 418 U/mg,纯度为85%.自制纤维素亲和膜有较好的吸附性能和纯化效果,重复使用性能良好,制备和纯化过程简便易行,具有广阔应用前景.     

高温胁迫对大豆幼苗生理特性的影响

以豫豆‘19’为材料,研究了大豆幼苗在适温(25℃)和高温(35℃,40℃)下叶片某些生理生化指标的变化,高温胁迫在人工气候箱中进行.结果表明:在高温下,以25℃为对照,随着胁迫时间的延长,RWC含量和总叶绿素(Chl)含量明显下降;MDA含量明显增加,膜脂过氧化作用加强;Pro含量也逐渐增加;SOD活性在胁迫初期明显增加,说明大豆幼苗可通过增加脯氨酸含量以及SOD活性来提高其耐热性.随着胁迫时间延长,SOD活性出现下降的趋势,大豆幼苗受害明显加重.相关分析表明,相对含水量(RWC)、丙二醛(MDA)和脯氨酸(Pro)含量呈显著负相关(r>-0.91),丙二醛(MDA)含量与脯氨酸(Pro)含量呈极显著相关(r>0.93),可以作为耐热性鉴定指标.     

干旱及外源SOD对盆栽莴苣抗旱指标的影响

以盆栽莴苣为试材,研究了干旱及外源超氧化物歧化酶(SOD)莴苣抗旱生理指标的影响。结果表明:在干旱胁迫下,莴苣SOD,过氧化物酶(POD)活性均呈现出先上升后下降的变化趋势,施用外源SOD延缓了其SOD活性的上升同时降低了POD上升的幅度;干旱使丙二醛(MDA)和脯氨酸(Pro)含量升高,施用外源SOD使严重干旱下两者上升强度得到缓解;干旱使叶绿素a与叶绿素b的含量比值增大,外源SOD使其下降并接近对照水平,说明外源SOD可能缓解了叶绿素b的降解;干旱使莴苣叶片水势早期下降,外源SOD使其下降过程延迟。说明外源SOD缓解了莴苣的干旱胁迫,使多项抗旱生理指标得到了改善,可以提高莴苣的抗旱性能。     

Salt and pH-dependent properties of native and mutant insulin

A fast and effective model for predicting the salt and pH dependent properties of protein complexes is presented. It is based on the formal charge parameter sets of ionizable groups and applied in conjunction with the finite difference Poisson-Boltzmann (FDPB) method to calculate the electrostatic interactions. All simulations were performed on the native 2Zn insulin and its fast-acting mutants such as B9D (B9Ser→ Asp), B9E (B9Aer→ Giu), B9EB10D (B9Ser→Glu, B10His→ Asp), and B10D (B10His → Asp). The salt and pH dependent properties of these dimers were analyzed from the aspect of electrostatic interaction, and the theoretical basis of the fast-acting behavior of these mutants was explained. It is found that the results agree well with experimental observations.     

Efficient inactivation of Staphylococcus aureus by silver and copper loaded photocatalytic titanate nanotubes

One dimensional titanate nanotubes(TNTs) were synthesized by microwave assisted alkaline hydrothermal process. The process was followed by UV-photodeposition of Ag and Cu on the surface of TNTs to enhance the photocatalytic activity in visible light spectrum. The loading of Ag and Cu(single and combination mode)offered a new insight to inactivate multi-drug resistant micro-organisms. The antibacterial properties of these samples were studied on Gram positive bacteria, Staphylococcus aureus(S. aureus) using well diffusion method.The TNTs with Ag and Cu loading showed a clear zone of inhibition after overnight incubation of S. aureus. The bacterial inactivation efficiency of nanoparticles in the visible light was further analyzed by kill kinetics. TNTs with Ag and/or Cu loading showed a significant reduction in bacterial growth. Cu co-loaded with Ag sample showed the highest inactivation efficiency within 90 min of visible light irradiation. To elucidate the mechanism of bactericidal properties of samples under visible light irradiation, the formation of reactive oxygen species(ROS), particularly, superoxide radical anion was determined by nitro blue tetrazolium(NBT) assay and the protein degradation by each samples were measured. Based on overall results, it was observed that the Cu coloaded with Ag on TNTs samples were found to be more effective as compared to either Ag or Cu loaded TNTs. It provides new avenues for utilizing the combination of Cu and Ag for enhancing the antimicrobial efficacies for different nanoparticles.     

中华猕猴桃营养生物学效应研究——Ⅲ.浓缩果汁对家兔血液红细胞的邻苯三酚自氧化抑制效应

采用邻苯三酚自氧化抑制法,对口服中华猕猴桃浓缩果汁家兔血液红细胞的邻苯三酚自氧化抑制效应的试验表明,按50kg体重人体,每日食用0.5kg果汁剂量,持续5周服用中华猕猴桃浓缩果汁的家免,其血液红细胞邻苯三酚自氧化反应抑制率明显升高。与服用前正常值73.7％对比,1—5周依次为129.1％,153.1％,133.4％,154.9％和150.8％。停服果汁2周和4周的测定值为111.8％和114.8％。结果提示,口服中华猕猴桃浓缩果汁有明显增强家兔血液红细胞的邻苯三酚自氧化抑制效应,抑制率比正常值提高1倍。猕猴桃果汁与同等剂量试剂抗坏血酸平行抑制试验结果,猕猴桃果汁的自氧化抑制率较试剂抗坏血酸高2个剂量级。     

Cu/Al管气体火焰钎焊接头特征及热力学分析

通过计算Cu/Al管氧乙炔气体火焰钎焊条件下形成金属间化合物的各化学反应的熵变,对Cu/Al金属间化合物的形成及向CuAl2转化的趋势进行了化学热力学分析;结合XRD、SEM、EDS研究了Cu/Al管氧乙炔气体火焰钎焊接头组织与元素分布特征。结果表明,Cu/Al管氧乙炔气体火焰钎焊条件下,接头中脆性金属间化合物CuAl2由Cu、Al原子的直接结合和其他Cu/Al金属间化合物与Al原子的继续反应生成,其中CuAl自主转化趋势较强;热力学计算分析与接头XRD分析结果一致。钎焊接头可分为3个特征区域:靠近Al基体侧形成了宽度约30μm的α-Al与α-Al+CuAl2二元共晶区;钎缝中心偏Al基体一侧形成了宽度约150μm组织细密的多元共晶组织区;钎缝中靠近Cu基体宽度约120μm区域,Cu的大量扩散并与Al充分反应,形成了粗大珊瑚状CuAl2。     

Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor

Endothelium-derived vascular relaxing factor (EDRF) is a humoral agent that is released by vascular endothelium and mediates vasodilator responses induced by various substances including acetylcholine and bradykinin. EDRF is very unstable, with a half-life of between 6 and 50 s, and is clearly distinguishable from prostacyclin. The chemical structure of EDRF is unknown but it has been suggested that it is either a hydroperoxy- or free radical-derivative of arachidonic acid or an unstable aldehyde, ketone or lactone. We have examined the role of superoxide anion (O-2) in the inactivation of EDRF released from vascular endothelial cells cultured on microcarrier beads and bioassayed using a cascade of superfused aortic smooth muscle strips. With this system, we have now demonstrated that EDRF is protected from breakdown by superoxide dismutase (SOD) and Cu2+, but not by catalase, and is inactivated by Fe2+. These findings indicate that O-2 contributes significantly to the instability of EDRF.