Structure of fumarate reductase from Wolinella succinogenes at 2.2 A resolution

Fumarate reductase couples the reduction of fumarate to succinate to the oxidation of quinol to quinone, in a reaction opposite to that catalysed by the related complex II of the respiratory chain (succinate dehydrogenase). Here we describe the crystal structure at 2.2 A resolution of the three protein subunits containing fumarate reductase from the anaerobic bacterium Wolinella succinogenes. Subunit A contains the site of fumarate reduction and a covalently bound flavin adenine dinucleotide prosthetic group. Subunit B contains three iron-sulphur centres. The menaquinol-oxidizing subunit C consists of five membrane-spanning, primarily helical segments and binds two haem b molecules. On the basis of the structure, we propose a pathway of electron transfer from the dihaem cytochrome b to the site of fumarate reduction and a mechanism of fumarate reduction. The relative orientations of the soluble and membrane-embedded subunits of succinate:quinone oxidoreductases appear to be unique.     

Reversible redox energy coupling in electron transfer chains

Reversibility is a common theme in respiratory and photosynthetic systems that couple electron transfer with a transmembrane proton gradient driving ATP production. This includes the intensely studied cytochrome bc1, which catalyses electron transfer between quinone and cytochrome c. To understand how efficient reversible energy coupling works, here we have progressively inactivated individual cofactors comprising cytochrome bc1. We have resolved millisecond reversibility in all electron-tunnelling steps and coupled proton exchanges, including charge-separating hydroquinone-quinone catalysis at the Q(o) site, which shows that redox equilibria are relevant on a catalytic timescale. Such rapid reversibility renders popular models based on a semiquinone in Q(o) site catalysis prone to short-circuit failure. Two mechanisms allow reversible function and safely relegate short-circuits to long-distance electron tunnelling on a timescale of seconds: conformational gating of semiquinone for both forward and reverse electron transfer, or concerted two-electron quinone redox chemistry that avoids the semiquinone intermediate altogether.     

怎样把QB程序快速转换为VB程序

本文介绍了把ＱＢ程序转换为ＶＢ程序的方法并且给出了一个实例。     

NaCl胁迫对椒样薄荷光合作用和PSII光化学活性的影响

利用光合作用测定仪和叶绿素荧光仪研究了不同浓度NaCl胁迫对椒样薄荷光合作用和PSII光化学活性的影响。结果表明,在NaCl胁迫下,椒样薄荷的生长和生物量呈现出了浓度依赖性的变化。在100和150 mmol/L NaCl胁迫下,椒样薄荷能够正常生长。随着NaCl处理浓度的提高,椒样薄荷光合能力和PSII光化学活性显著下降,表明QA到QB之间的电子传递被阻断。光合作用对NaCl胁迫下植物生长和代谢的影响最为突出,PSII是光合系统中最敏感的组分,在植物光合系统对环境胁迫的应激中起着重要的作用。     

Novel thioether bond revealed by a 1.7 A crystal structure of galactose oxidase

Galactose oxidase is an extracellular enzyme secreted by the fungus Dactylium dendroides. It is monomeric, with a relative molecular mass of 68,000, catalyses the stereospecific oxidation of a broad range of primary alcohol substrates and possesses a unique mononuclear copper site essential for catalysing a two-electron transfer reaction during the oxidation of primary alcohols to corresponding aldehydes. Recent evidence arguing against a Cu(III)-Cu(I) couple implies the existence of a second redox-active site proposed to involve pyrroloquinoline quinone or a tyrosine radical. We now report the crystal structure of galactose oxidase at 1.7 A resolution. This reveals a unique structural feature at the copper site with a novel thioether bond linking Cys 228 and Tyr 272 in a stacking interaction with Trp 290. We propose that these molecular components stabilize the protein free-radical species essential for catalysis and thus provide a 'built-in' secondary cofactor. This feature may represent a new mechanism for mediating electron transfer in metalloenzymes in the absence of exogenous cofactors.     

框支-短肢剪力墙结构中斜柱转换结构的抗震试验研究

通过对两个单榀框支—短肢剪力墙斜柱转换结构在竖向荷载及水平荷载共同作用下的静力试验,分析了构件内应力的分布状态、试件的破坏形态、荷载传递以及转换梁的受力性能和构件的抗震性能。试验结果表明框支—短肢剪力墙斜柱转换结构具有较好的抗震性能。同时提出了在工程实践中该结构设计应注意的问题。     

Intraprotein radical transfer during photoactivation of DNA photolyase

Amino-acid radicals play key roles in many enzymatic reactions. Catalysis often involves transfer of a radical character within the protein, as in class I ribonucleotide reductase where radical transfer occurs over 35 A, from a tyrosyl radical to a cysteine. It is currently debated whether this kind of long-range transfer occurs by electron transfer, followed by proton release to create a neutral radical, or by H-atom transfer, that is, simultaneous transfer of electrons and protons. The latter mechanism avoids the energetic cost of charge formation in the low dielectric protein, but it is less robust to structural changes than is electron transfer. Available experimental data do not clearly discriminate between these proposals. We have studied the mechanism of photoactivation (light-induced reduction of the flavin adenine dinucleotide cofactor) of Escherichia coli DNA photolyase using time-resolved absorption spectroscopy. Here we show that the excited flavin adenine dinucleotide radical abstracts an electron from a nearby tryptophan in 30 ps. After subsequent electron transfer along a chain of three tryptophans, the most remote tryptophan (as a cation radical) releases a proton to the solvent in about 300 ns, showing that electron transfer occurs before proton dissociation. A similar process may take place in photolyase-like blue-light receptors.     

Directional electron transfer in ruthenium-modified horse heart cytochrome c

Cytochrome c can be modified by [(NH3)5RuII/III-] specifically at the imidazole moiety of histidine 33, and we have recently discussed the thermodynamics and kinetics of electron transfer within this modified protein. X-ray crystal structures of the oxidized and reduced forms of tuna cytochrome c indicate that the separation between the haem group of cytochrome c and the ruthenium label is 12-16 A. Internal electron transfer from the [(NH3)5RuII-] centre to the Fe(III) haem centre occurs with a rate constant k congruent to 53 s-1 (25 degrees C) (delta H = 3.5 kcal mol-1, delta S = -39 EU), as measured by pulse radiolysis. The measured unimolecular rate constant, k congruent to 53 s-1, is on the same timescale as a number of conformational changes that occur within the cytochrome c molecule. These results raise the question of whether electron transfer or protein conformational change is the rate limiting step in this process. We describe here an experiment that probes this intramolecular electron transfer step further. It involves reversing the direction of electron transfer by changing the redox potential of the ruthenium label. Electron transfer in the new ruthenium-cytochrome c derivative described here is from haem(II) to the Ru(III) label, whereas in (NH3)5Ru-cytochrome c the electron transfer is from Ru(II) to haem(III). Intramolecular electron transfer from haem(II) to Ru(III) in the new ruthenium-cytochrome c described here proceeds much slower (greater than 10(5) times) than the electron transfer from Ru(II) to haem(III) in the (NH3)5Ru-cytochrome c. We therefore conclude that electron transfer in cytochrome c is directional, with the protein envelope presumably involved in this directionality.     

应用巴特沃思逼近实现三阶有源低通滤波器的设计

本文介绍的设计,是应用巴特沃思逼近找到一个可实现的三阶电压转移函数,并用RC一阶有源电路和RC二阶有源电路的级联,来获得能满足上述电压转移函数的三阶有源低通滤波器。文中还提出了在设计中采用归一化方法的好处,以及去归一化的方法。     

梁侧锚固钢板加固混凝土梁的横向剪力传递模型

为研究梁侧锚固钢板加固钢筋混凝土梁(BSP梁)中钢-混凝土连接界面上的横向滑移对加固梁的极限承载力及变形性能的影响,将BSP梁中横向剪力传递类比于Winkler弹性地基模型,并结合已有试验和数值模拟成果,提出了可用于计算横向滑移和横向剪力传递的分段线性简化模型.从而得出了由混凝土梁和钢板抗弯刚度及螺栓连接剪切刚度计算横向滑移和横向剪力传递的实用计算方法.该简化模型的适用性得到了试验成果的检验,并可用于指导BSP梁的加固设计.     

RC二端口网络电压传递函数的研究

本文从RC二端口网络的电压传递函数及入端导纳的表达式出发,研究了电压传递函数,入端导纳,负载导纳之间的关系以及它们和网络Y参数的关系,得出了几个重要关系式。这些结论对丰富RC二端口网络的综合方法提供了必要的理论基础。     

Nature of biological electron transfer.

Powerful first-order analysis of intraprotein electron transfer is developed from electron-transfer measurements both in biological and in chemical systems. A variation of 20 A in the distance between donors and acceptors in protein changes the electron-transfer rate by 10(12)-fold. Protein presents a uniform electronic barrier to electron tunnelling and a uniform nuclear characteristic frequency, properties similar to an organic glass. Selection of distance, free energy and reorganization energy are sufficient to define rate and directional specificity of biological electron transfer, meeting physiological requirements in diverse systems.     

SRC—RC 竖向混合结构转换柱研究现状综述

通过16根转换柱试件及钢筋混凝土柱对比试件的低周反复荷载试验,分析型钢延伸高度、箍筋配置和型钢配钢率对转换柱受力性能的影响,为SRC-RC竖向混合结构的推广应用提供科研基础与参考.指出:(a)由于型钢的局部存在,转换柱容易产生类似于RC短柱的剪切破坏,破坏主要集中在RC部分,钢与混凝土之间的共同工作会导致此类破坏发生;(b)为了保证转换柱具有更好的抗震性能,应采取构造措施控制剪切裂缝的发展,避免剪切破坏,提高构件的变形能力.建议构件全高箍筋加密或在RC部分设置X形交叉钢筋.在对转换柱特殊破坏方式充分认知的基础上,对新的构造措施与配钢方式进行了展望.     

组合式钢框架内填预制RC墙结构静力性能有限元分析

钢框架内填预制钢筋混凝土剪力墙结构是新型混合式结构,由钢框架与内填墙组成双重防线,具有良好的抗侧力能力,同时预制构配件和预制装配建筑有利于推动住宅产业化发展.考虑大尺寸内填预制RC墙运输和安装困难的情况,提出竖向和横向组合式钢框架内填RC墙结构,采用ABAQUS建立有限元模型进行结构受力分析.通过分析荷载位移曲线,构件应力分布和变形情况,研究结构破坏特点和受力性能.结果表明,全螺栓结构因其合理的传力路径,有良好的承载力和延性;竖向组合式具有较好的初始刚度和整体承载力,与全螺栓连接预制RC墙有近似的受力性能,便于运输和安装;而横向组合式由于上下板缺乏有效传力路径,初始刚度和最终承载力都明显低于全螺栓和竖向组合式,不利于实际工程应用.     

Electrostatic effect on electron transfer between cytochrome b5 and cytochrome c

The binding and electron transfer between wild type, E44A, E56A, E44/56A, E44/48/56A/D60A and F35Y variants of cytochrome b5 and cytochrome c were studied. When mixed with cytochrome c, the cytochrome b, E44/48/56A/D60A did not show the typical UV-vis difference spectrum of absorption, indicating that the alteration of the surface electrostatic potential obviously influenced the spectrum. The electron transfer rates of wild type cytochrome bj, its variants and cytochrome c at different temperature and ionic strength exhibited an order of F35Y > wild type > E56A > E44A > E44/48/56A/D60A. The enthalpy and entropy of the reaction did not change obviously, suggesting that the mutation did not significantly disturb the electron transfer conformation. The investigation of electron transfer rate constants at different ionic strength demonstrated that electrostatic interaction obviously affected the electron transfer process. The significant difference of Cyt b, F35Y and E44/48/56A/D60A from the wild type protein further confirmed the great importance of the electrostatic interaction in the protein electron transfer.     

Electrostatic effect on electron transfer between cytochrome b5 and cytochrome c

The binding and electron transfer between wild type, E44A, E56A, E44/56A, E44/48/56A/D60Aand F35Y variants of cytochrome b5 and cytochrome c were studied. When mixed with cytochrome c, the cytochrome b5E44/48/56A/D60A did not show the typical UV-vis difference spectrum of absorption, indicating that the alteration ofthe surface electrostatic potential obviously influenced the spectrum. The electron transfer rates of wild type cytochromeb5, its variants and cytochrome e at different temperature and ionic strength exhibited an order of F35Y > wild type >E56A > E44A > E44/48/56A/D60A. The enthalpy and entropy of the reaction did not change obviously, suggestingthat the mutation did not significantly disturb the electron transfer conformation. The investigation of electron transfer rateconstants at different ionic strength demonstrated that electrostatic interaction obviously affected the electron transfer pro-cess. The significant difference of Cyt b5 F35Y and E44/48/56A/D60A from the wild type protein further confirmed thegreat importance of the electrostatic interaction in the protein electron transfer.     

短小芽孢杆菌质粒pCJ转化枯草杆菌的研究

本文采用我国自己分离的短小芽孢杆菌抗四环素质粒pCJ3转化枯草杆菌的各种突变体.质粒DNA经氯化铯—溴化乙锭密度梯度离心纯化后转化枯草杆菌的感受态细胞.结果表明枯草杆菌BR151,SB202,168,QB1130及QB1133都可作为质粒pCJ3的受体,转化效率在10~3左右,其中以SB202这株突变体为最高,从各种转化体中提取的质粒及经BamHI消化后的质粒的电泳图均与原质粒pCJ3及其BamHI酶切片段相同,并具有pCJ3的抗四环素转化活性.将pCJ3DNA经BamHI酶切后重新用T_4-DNA连接酶连接再转化枯草杆菌细胞,其转化效率比原质粒高1—2个数量级.研究了二价金属离子、pH及培养基成分对转化的影响,结果证明:Ca~(++)对枯草杆菌转化有促进作用,Cu~(++),Zn~(++)则有抑制作用,转化的最适pH在7.2—7.5之间;营养丰富培养能提高转化效率.     

An atypical haem in the cytochrome b(6)f complex

Photosystems I and II (PSI and II) are reaction centres that capture light energy in order to drive oxygenic photosynthesis; however, they can only do so by interacting with the multisubunit cytochrome b(6)f complex. This complex receives electrons from PSII and passes them to PSI, pumping protons across the membrane and powering the Q-cycle. Unlike the mitochondrial and bacterial homologue cytochrome bc(1), cytochrome b(6)f can switch to a cyclic mode of electron transfer around PSI using an unknown pathway. Here we present the X-ray structure at 3.1 A of cytochrome b(6)f from the alga Chlamydomonas reinhardtii. The structure bears similarities to cytochrome bc(1) but also exhibits some unique features, such as binding chlorophyll, beta-carotene and an unexpected haem sharing a quinone site. This haem is atypical as it is covalently bound by one thioether linkage and has no axial amino acid ligand. This haem may be the missing link in oxygenic photosynthesis.     

采用ANSYS分析方柱-T形薄壁柱局部转换节点的受力特征

文中采用有限元分析程序ANSYS对三个钢筋混凝土方柱-T形薄壁柱局部转换节点在竖向荷载作用下的受力性能进行分析,得出了转换梁的加腋高度不仅影响自身的承载能力,还对其上的薄壁柱的应力状态有直接影响的结论.展示了有限元分析应用于工程设计内力计算的方法.     

QB DNA-containing hybrid plasmids giving rise to QB phage formation in the bacterial host.

Hybrid plasmids consisting of PCRI and a complete DNA copy of the RNA genome of coli.phage QB, inserted in either orientation, elicit the formation of phage QB when introduced into Escherichia coli