Architecture of succinate dehydrogenase and reactive oxygen species generation

The structure of Escherichia coli succinate dehydrogenase (SQR), analogous to the mitochondrial respiratory complex II, has been determined, revealing the electron transport pathway from the electron donor, succinate, to the terminal electron acceptor, ubiquinone. It was found that the SQR redox centers are arranged in a manner that aids the prevention of reactive oxygen species (ROS) formation at the flavin adenine dinucleotide. This is likely to be the main reason SQR is expressed during aerobic respiration rather than the related enzyme fumarate reductase, which produces high levels of ROS. Furthermore, symptoms of genetic disorders associated with mitochondrial SQR mutations may be a result of ROS formation resulting from impaired electron transport in the enzyme.     

不同铵硝比对菠菜有机酸和淀粉含量的影响

【目的】研究不同铵硝比的氮素营养对于菠菜有机酸和淀粉含量的影响。【方法】采用营养液栽培菠菜的方法,营养液中的氮素形态设置硝态氮和铵态氮以一定比例（100﹕0、75﹕25、50﹕50、25﹕75 和 0﹕100）配合。【结果】随着铵硝比的下降,（1）菠菜茎叶丙酮酸、柠檬酸、α-酮戊二酸、琥珀酸、延胡索酸和苹果酸等6种有机酸含量以及淀粉含量都呈上升趋势;同时,有机酸含量与营养液中初始硝态氮浓度之间呈显著的线性正相关关系;（2）菠菜根系丙酮酸、柠檬酸、琥珀酸和苹果酸含量以及淀粉含量也呈上升趋势,但延胡索酸含量的变化不明显;（3）除了柠檬酸外,菠菜根系其它5种有机酸含量与其茎叶中相应有机酸含量的比值都呈现下降的趋势;在全硝营养条件下,根系中只有琥珀酸和延胡索酸的含量低于其在茎叶中的含量;而在全铵营养时,菠菜根系中上述5种有机酸的含量均明显高于茎叶中的相应有机酸的含量。因此,与根系相比,菠菜茎叶中5种有机酸含量随营养液中硝态氮比例增加而增加的幅度更明显。而随着营养液中硝态氮比例的增加,菠菜根系柠檬酸含量与其茎叶柠檬酸含量的比值逐渐升高,则说明根系柠檬酸含量增加的幅度要大于茎叶柠檬酸含量;（4）菠菜的茎叶和根系中的淀粉含量都呈现下降的趋势,且与营养液中硝态氮浓度之间均呈现为显著的负相关关系。【结论】随着营养液中硝态氮比例的增加,菠菜茎叶和根系有机酸代谢均表现为增强的趋势。     

植物组织中有机酸的提取方法比较

以番茄果实和不结球白菜叶片为试验材料,用蒸馏水（水提）、80％乙醇-旋转蒸发浓缩（醇提）、蒸馏水-冷冻干燥浓缩（水提冻干）分别提取有机酸,采用高效液相色谱法测定。结果表明：3种提取方法下番茄果实中富马酸、丁二酸含量无显著性差异,草酸、酒石酸、苹果酸、草酰乙酸含量均以水提最高,而丙酮酸和柠檬酸以醇提含量最高;不同提取方法对不结球白菜叶片中草酸、酒石酸、富马酸含量无影响,其他几种有机酸（除草酰乙酸外）均以水提和水提冻干含量高于醇提,其中,水提法和水提冻干法提取丙酮酸、苹果酸、丁二酸的含量无显著差异,而柠檬酸含量以水提冻干法最高。综合考虑试验步骤和结果等各方面因素发现,蒸馏水直接提取法既能有效保证有机酸的提取量,又能大大简化试验步骤。     

Significance of mitochondria for porphyrin and heme biosynthesis

It is concluded that mitochondria are involved in three steps of porphyrin and heme biosynthesis-first, in the formation of delta-aminolevulinic acid from glycine and active succinate; second, in the synthesis of protoporphyrin; third, in the incorporation of iron into the porphyrin ring-that is, in heme formation.     

Regulation of the erythropoietin gene: evidence that the oxygen sensor is a heme protein

Erythropoietin (Epo), the hormone that stimulates red blood cell production, is synthesized in the kidney and liver in response to hypoxia. The human hepatoma cell line Hep3B regulates its production of Epo in a physiologic manner. Either hypoxia or cobalt chloride markedly increases expression of Epo mRNA as well as production of biologically active and immunologically distinct Epo protein. New protein synthesis is required before the induction of increased levels of hypoxia- or cobalt-induced Epo mRNA. Hypoxia, cobalt chloride, and nickel chloride appear to stimulate Epo production through a common pathway. The inhibition of Epo production at low partial pressures of oxygen by carbon monoxide provides evidence that a heme protein is integrally involved in the oxygen-sensing mechanism. This hypothesis is further supported by the finding that when heme synthesis is blocked, hypoxia-, cobalt-, and nickel-induced Epo production are all markedly inhibited. A model is proposed in which a ligand-dependent conformational change in a heme protein accounts for the mechanism by which hypoxia as well as cobalt and nickel stimulate the production of Epo.     

Evidence for an inter-organismic heme biosynthetic pathway in symbiotic soybean root nodules

The successful symbiosis of soybean with Bradyrhizobium japonicum depends on their complex interactions, culminating in the development and maintenance of root nodules. A B. japonicum mutant defective in heme synthesis in culture was able to produce heme as a result of its symbiotic association with the soybean host. The bacterial mutant was incapable of synthesizing the committed heme precursor delta-aminolevulinic acid (ALA), but nodule plant cells formed ALA from glutamate. In addition, exogenous ALA was taken up by isolated nodule bacteria of the parent strain and of the mutant. It is proposed that bacterial heme found in nodules can be synthesized from plant ALA, hence segments of a single metabolic pathway are spatially separated into two organisms.     

Hyperbaric oxygen: effects on metabolism and ionic movement in cerebral cortex slices

The incubation of slices of cerebral cortex under hyperbaric oxygen pressures from 1 to 10 atmospheres in the presence of radioactive glucose, pyruvate, succinate, fumarate, L-glutamate, and gamma-aminobutyric acid causes a marked diminution of tissue oxidative reactions. There is a simultaneous decrease in phosphocreatine and adenosine triphosphate, and a reduction of the apparent intracellular ionic gradients. The increase of lipid peroxides, measured directly, is attributed to the toxic effects of hyperbaric oxygen.     

New light-sensitive cofactor required for oxidation of succinate by Mycobacterium phlei

Irradiation of the electron transport particles of Mycobacterium phlei with light at a wavelength of 360 manometers resulted in a loss of oxidase activities of succinate and the reduced form of nicotinamide adenine dinucleotide. The lesion in the two pathways caused by irradiation of the particles differs. The succinoxidase pathway was more labile to irradiation than the pathway linked to nicotinamide adenine dinucleotide. Restoration of succinoxidase activity (up to 50 to 60 percent) occurred on addition of a thermostable, water-soluble material obtained from Mycobacterium phlei cells or with an extract of mitochondria from boiled rat liver. Other known cofactors, such as flavine adenine dinucleotide, flavine mononucleotide, benzo- and naphthoquinones, as well as sulfhydryl agents, failed to restore succinoxidase activity after irradiation. Water-soluble material from Mycobacterium phlei appears to function between the flavoprotein and cytochrome b on the succinoxidase pathway. In contrast to the requirements for restoration of the pathway linked to nicotinamide adenine dinucleotide, restoration of succinoxidase does not occur with quinones or other cofactors such as flavine adenine dinucleotide.     

Shunt bilirubin: evidence for two components

Studies with C(14)-labeled glycine and delta-aminolevulinic acid as heme-bilirubin precursors in man indicate that the early labeled or shunt bilirubin consists of two fractions. Fraction 1 requires 1 to 24 hours for maximum synthesis, is not dependent on marrow erythropoietic heme synthesis, and is possibly of anabolic origin (formed by a direct pathway from heme precursors). Fraction 2 requires 3 to 4 days for maximum production, is dependent on heme synthesis, and probably has its origin in the bone marrow, as a degradation product of red-cell heme.     

超高效液相色谱-串联质谱法测定糕点中富马酸二甲酯

[目的]简便快捷地测定糕点中的防腐剂富马酸二甲酯。[方法]建立了一种使用岛津超高效液相色谱仪LC-30A和三重四级杆质谱仪API 4000联用测定食品中富马酸二甲酯含量的分析方法。[结果]试验表明,富马酸二甲酯标液的线性关系为y=0.972 7x+0.001 2,线性相关系数为0.999 4。采取加标的方式进行方法验证,对同一富马酸二甲酯浓度的加标样品进行6次精密度试验,富马酸二甲酯的回收率范围为98.4%～101.5%,相对标准偏差RSD为1.2%,方法检出限为0.01 mg/L。[结论]该方法准确、快速、操作简便,可用于糕点中富马酸二甲酯的定量分析。     

纳米铜对大鼠肝脏毒性相关蛋白过氧化氢酶的分离鉴定及生物信息学分析

【目的】分离和鉴定纳米铜对大鼠肝脏毒性相关蛋白过氧化氢酶(catalase, CAT),探讨CAT在毒性发挥中的作用,为揭示纳米铜对肝脏毒性机制提供依据。【方法】应用2-DE技术和PDQuest 8.0软件在大鼠肝脏蛋白组中筛选纳米铜对肝脏毒性差异蛋白,经质谱鉴定后进行生物信息学分析。【结果】筛选到下调的差异蛋白点6602和7702与肝毒性相关,鉴定均为CAT蛋白;其性质稳定,有一定亲水性,无信号肽,定位于细胞质,可能属于非分泌性蛋白,含有过氧化氢酶活性位点64FDRERIPERVVHAKGAG80和过氧化氢酶亚铁血红素配合基位点354RLFAYPDTH362等功能位点;无规则卷曲、α螺旋和延伸链是其主要的二级结构元件,并预测了其三级结构图;同源性分析表明,大鼠的CAT与其它8个物种有较高同源性,并构建了CAT 蛋白的系统进化树。【结论】纳米铜通过下调大鼠肝脏中CAT蛋白表达,引起肝细胞氧化应激损伤,可能是其发挥毒性作用的途径之一。     

Terminal oxidation in the regulation of heme biosynthesis

Turnover of the tricarboxylic acid cycle and the rate of succinyl-coenzyme A formation may be important factors in the regulation of heme biosynthesis in liver homogenate. Acting as hydrogen acceptor, acetoacetate appears to have a unique role in influencing these metabolic processes.     

赤豆种子萌发中酰脲累积动态及若干碳氮化合物对其形成的影响

赤豆种子萌发中,幼苗迅速合成酰脲,萌发第4d达到最大值.谷氨酰胺和甘氨酸促进尿囊素合成,谷氨酸和天冬酰胺对尿囊素合成影响较小.外源铵促进幼苗合成尿囊素.三羧酸循环的部分中间产物(特别是琥珀酸或延胡索酸)或高浓度的乙醛酸(6-10 mmol/L)不同程度地抑制幼苗的尿囊索合成.     

延胡索酸泰妙菌素可溶性粉对猪支原体肺炎的疗效研究

[目的]评价延胡索酸泰妙菌素可溶性粉对猪支原体肺炎的治疗效果。[方法]在水中添加30、45、60mg／L延胡索酸泰妙菌素可溶性粉,对猪支原体肺炎进行混饮治疗,研究延胡索酸泰妙菌素可溶性粉对猪支原体肺炎的治疗效果。[结果]与支原净对照组相比,延胡索酸泰妙菌素可溶性粉Ⅱ、Ⅲ有效率、治愈率、肺损伤改善率、相对日增重率、料肉比与支原净基本相同。延胡索酸泰妙菌素可溶性粉剂量为60mg／L试验组中有效率为100％,治愈率为95％。延胡索酸泰妙茵素可溶性粉剂量为45瑚唔／L试验组中有效率95％,治愈率为90％。[结论]延胡索酸泰妙菌素可溶性粉在饮水中的推荐混合荆量为45—60mg／L。     

反丁烯二酸桂醇甲酯的合成及抗菌活性研究

 以α,β-不饱和羰基结构为母体设计合成反丁烯二酸桂醇甲酯(CMF),并对反丁烯二酸桂醇甲酯的抗菌活性进行了研究。合成试验表明:以摩尔比为1∶2的马来酸酐与肉桂醇在95℃醇解反应2~4h后加入异构化催化剂,90℃转型反应4h后,添加6倍摩尔质量的甲醇,在90℃恒温反应4h,反丁烯二酸桂醇甲酯的合成产率较高。皮肤刺激性试验表明:CMF对皮肤无致敏性。抗菌活性试验表明:反丁烯二酸桂醇甲酯不但杀菌谱广,而且有效作用浓度低,在食品加工、保藏中具有广阔的开发应用前景。     

Bilirubin inhibition of heme biosynthesis

The conversion of protoporphyrin and iron to heme is catalyzed by a soluble enzyme prepared from rat liver. This reaction is inhibited by bilirubin, and initial kinetic studies suggest that the inhibition is due in part to a competition between protoporphyrin and the bile pigment. Implications of this finding in hyperbilirubinemia are mentioned.     

"Apocatalase" of catalase-negative staphylococci

Crystalline catalase of high purity and activity was prepared from a heme-requiring Staphylococcus aureus mutant, JT/52, grown in the presence of heme. A crystalline protein was obtained from the same cells grown without heme. This enzymatically inactive protein and the catalase cross-react strongly when tested by a variety of immunochemical tests.     

小鼠肝再生过程中脂质代谢相关通路中基因的表达谱变化

[目的]研究肝再生过程中肝脏内脂质代谢通路相关基因的表达谱变化。[方法]建立CC l4诱导肝再生小鼠模型,从小鼠肝组织中提取总RNA,通过微阵列基因芯片技术检测在不同肝再生期间脂质代谢通路相关基因的表达变化,并进行具体功能分析。[结果]肝再生过程中,有98个脂质代谢相关基因的表达水平发生了变化,根据这些基因表达的变化趋势可以分为8组。整体上看,基因表达前期表现为抑制,后期表现为上调。其中,脂肪酸的合成通路基因表达以上调为主,分解代谢通路变化并不明显;大多数胆汁酸合成通路基因在4.5天之前表现为抑制,在4.5天和7天时表现为上调。[结论]肝再生过程中,脂质代谢相关通路间的基因表达变化并不一致,这些数据为进一步研究脂质代谢相关通路对肝再生的调控作用提供了明确的基因范围。     

小鼠肝再生过程中脂质代谢相关通路中基因的表达谱变化

1材料与方法 1．1材料 试验动物：9周龄的雄性C57BIV6小鼠，自由采食和饮水，每天光照12h，黑暗12h。试剂：TRIzol试剂（Invitrogen）；RNeasy Total RNA Mini Kit（Qiagen）；基因芯片Genechip mouse genome 43020（Affymetrix）；17启动子引物序列（Affymetrix）。     

Heme requirement for reproduction of a free-living nematode

The free-living hermaphroditic nematode Caernorhabditis briggsae has a nutritional requirement for heme. The organism can be subcultured repeatedly in a chemically defined axenic medium that contains autoclaved bacterial cells (Escherichia coli) and sterols if a hemeprotein-containing fraction from liver is present. Pure myoglobin, hemoglobin, cytochrome c, and hemin, respectively, can substitute effectively for the liver fraction.