Crystal structure of the sodium-potassium pump

The Na+,K+-ATPase generates electrochemical gradients for sodium and potassium that are vital to animal cells, exchanging three sodium ions for two potassium ions across the plasma membrane during each cycle of ATP hydrolysis. Here we present the X-ray crystal structure at 3.5 A resolution of the pig renal Na+,K+-ATPase with two rubidium ions bound (as potassium congeners) in an occluded state in the transmembrane part of the alpha-subunit. Several of the residues forming the cavity for rubidium/potassium occlusion in the Na+,K+-ATPase are homologous to those binding calcium in the Ca2+-ATPase of sarco(endo)plasmic reticulum. The beta- and gamma-subunits specific to the Na+,K+-ATPase are associated with transmembrane helices alphaM7/alphaM10 and alphaM9, respectively. The gamma-subunit corresponds to a fragment of the V-type ATPase c subunit. The carboxy terminus of the alpha-subunit is contained within a pocket between transmembrane helices and seems to be a novel regulatory element controlling sodium affinity, possibly influenced by the membrane potential.     

Crystal structure of the calcium pump with a bound ATP analogue

P-type ATPases are ATP-powered ion pumps that establish ion concentration gradients across cell and organelle membranes. Here, we describe the crystal structure of the Ca2+ pump of skeletal muscle sarcoplasmic reticulum, a representative member of the P-type ATPase superfamily, with an ATP analogue, a Mg2+ and two Ca2+ ions in the respective binding sites. In this state, the ATP analogue reorganizes the three cytoplasmic domains (A, N and P), which are widely separated without nucleotide, by directly bridging the N and P domains. The structure of the P-domain itself is altered by the binding of the ATP analogue and Mg2+. As a result, the A-domain is tilted so that one of the transmembrane helices moves to lock the cytoplasmic gate of the transmembrane Ca2+-binding sites. This appears to be the mechanism for occluding the bound Ca2+ ions, before releasing them into the lumen of the sarcoplasmic reticulum.     

Crystal structure of the calcium pump of sarcoplasmic reticulum at 2.6 A resolution

Calcium ATPase is a member of the P-type ATPases that transport ions across the membrane against a concentration gradient. Here we have solved the crystal structure of the calcium ATPase of skeletal muscle sarcoplasmic reticulum (SERCA1a) at 2.6 A resolution with two calcium ions bound in the transmembrane domain, which comprises ten alpha-helices. The two calcium ions are located side by side and are surrounded by four transmembrane helices, two of which are unwound for efficient coordination geometry. The cytoplasmic region consists of three well separated domains, with the phosphorylation site in the central catalytic domain and the adenosine-binding site on another domain. The phosphorylation domain has the same fold as haloacid dehalogenase. Comparison with a low-resolution electron density map of the enzyme in the absence of calcium and with biochemical data suggests that large domain movements take place during active transport.     

Crystal structure of a human membrane protein involved in cysteinyl leukotriene biosynthesis

The cysteinyl leukotrienes, namely leukotriene (LT)C4 and its metabolites LTD4 and LTE4, the components of slow-reacting substance of anaphylaxis, are lipid mediators of smooth muscle constriction and inflammation, particularly implicated in bronchial asthma. LTC4 synthase (LTC4S), the pivotal enzyme for the biosynthesis of LTC4 (ref. 10), is an 18-kDa integral nuclear membrane protein that belongs to a superfamily of membrane-associated proteins in eicosanoid and glutathione metabolism that includes 5-lipoxygenase-activating protein, microsomal glutathione S-transferases (MGSTs), and microsomal prostaglandin E synthase 1 (ref. 13). LTC4S conjugates glutathione to LTA4, the endogenous substrate derived from arachidonic acid through the 5-lipoxygenase pathway. In contrast with MGST2 and MGST3 (refs 15, 16), LTC4S does not conjugate glutathione to xenobiotics. Here we show the atomic structure of human LTC4S in a complex with glutathione at 3.3 A resolution by X-ray crystallography and provide insights into the high substrate specificity for glutathione and LTA4 that distinguishes LTC4S from other MGSTs. The LTC4S monomer has four transmembrane alpha-helices and forms a threefold symmetric trimer as a unit with functional domains across each interface. Glutathione resides in a U-shaped conformation within an interface between adjacent monomers, and this binding is stabilized by a loop structure at the top of the interface. LTA4 would fit into the interface so that Arg 104 of one monomer activates glutathione to provide the thiolate anion that attacks C6 of LTA4 to form a thioether bond, and Arg 31 in the neighbouring monomer donates a proton to form a hydroxyl group at C5, resulting in 5(S)-hydroxy-6(R)-S-glutathionyl-7,9-trans-11,14-cis-eicosatetraenoic acid (LTC4). These findings provide a structural basis for the development of LTC4S inhibitors for a proinflammatory pathway mediated by three cysteinyl leukotriene ligands whose stability and potency are different and by multiple cysteinyl leukotriene receptors whose functions may be non-redundant.     

Crystal structure of the bacterial membrane protein TolC central to multidrug efflux and protein export

Diverse molecules, from small antibacterial drugs to large protein toxins, are exported directly across both cell membranes of gram-negative bacteria. This export is brought about by the reversible interaction of substrate-specific inner-membrane proteins with an outer-membrane protein of the TolC family, thus bypassing the intervening periplasm. Here we report the 2.1-A crystal structure of TolC from Escherichia coli, revealing a distinctive and previously unknown fold. Three TolC protomers assemble to form a continuous, solvent-accessible conduit--a 'channel-tunnel' over 140 A long that spans both the outer membrane and periplasmic space. The periplasmic or proximal end of the tunnel is sealed by sets of coiled helices. We suggest these could be untwisted by an allosteric mechanism, mediated by protein-protein interactions, to open the tunnel. The structure provides an explanation of how the cell cytosol is connected to the external environment during export, and suggests a general mechanism for the action of bacterial efflux pumps.     

Proton-coupled electron transfer drives the proton pump of cytochrome c oxidase

Electron transfer in cell respiration is coupled to proton translocation across mitochondrial and bacterial membranes, which is a primary event of biological energy transduction. The resulting electrochemical proton gradient is used to power energy-requiring reactions, such as ATP synthesis. Cytochrome c oxidase is a key component of the respiratory chain, which harnesses dioxygen as a sink for electrons and links O2 reduction to proton pumping. Electrons from cytochrome c are transferred sequentially to the O2 reduction site of cytochrome c oxidase via two other metal centres, Cu(A) and haem a, and this is coupled to vectorial proton transfer across the membrane by a hitherto unknown mechanism. On the basis of the kinetics of proton uptake and release on the two aqueous sides of the membrane, it was recently suggested that proton pumping by cytochrome c oxidase is not mechanistically coupled to internal electron transfer. Here we have monitored translocation of electrical charge equivalents as well as electron transfer within cytochrome c oxidase in real time. The results show that electron transfer from haem a to the O2 reduction site initiates the proton pump mechanism by being kinetically linked to an internal vectorial proton transfer. This reaction drives the proton pump and occurs before relaxation steps in which protons are taken up from the aqueous space on one side of the membrane and released on the other.     

Experimental measurement of proton conductivity and electronic conductivity of membrane electrode assembly for proton exchange membrane fuel cells

Charge transport processes involving the proton migration and electron transfer for different parts of membrane electrode assemble (MEA) play an essential role for developing the novel electrode and enhancing the electrochemical performance towards proton exchange membrane fuel cells (PEMFCs). However, the coupled charge transport processes make it difficult for evaluating proton conductivity and electronic conductivity of different parts in MEAs under operation conditions of fuel cells. Here in this work, we propose an experiment approach for separating the electronic conductivity and proton conductivity of different components of MEA at the operating conditions of PEMFCs. This approach involves two different measuring devices, which both consist of electron or proton conducting layers, sealing layers and sample layer, followed by tailoring the thickness of sample layers and via electrochemical impedance spectroscopy (EIS) to quantity the electronic conductivity and proton conductivity of different layers. These experiment results show the great potential in the development of different components of MEA.     

环丙沙星晶体结构

在水热条件下,以Co(NO3)·6H2O、环丙沙星、对羟基苯甲酸为反应物,得到了晶体(C41H44F2N8O15).晶体的晶胞参数为a=10.0226(4)(A),b=13.4560(5)(A),c=16.3134(6)(A),α=98.030(2)°β=92.944(2)°,γ=111.008(2)°,V=2021.4(2)A3,Z=2,Dc=1.523 mg/m3.     

Crystal structure of leucine-enkephalin

The crystal structure of leucine-enkephalin has been determined in a crystal form that has four independent enkephalin molecules and much water and dimethylformamide solvent in the asymmetric unit. All four enkephalins have extended peptide backbones with the Tyr, Phe and Leu side chains above and below the plane of the backbone. There is evidence that this extended conformation may provide an acceptable model for enkephalin binding to opiate mu-receptors.     

质子交换膜燃料电池分体式集成电堆

质子交换膜燃料电池(PEMFC)是电动汽车的最佳候选电源。为提高其效率,研制了一种分体式质子交换膜燃料电池集成电堆。采用外置式Nafion○R115膜加湿器,加湿系统与冷却系统合二为一。在70℃,气体流量小于7L/min时,膜加湿器能为反应气体提供摩尔分数为72%的加湿量,满足了电堆的加湿需求。电堆活性面积为390cm2,在电流密度为2.1A/cm2时,电堆最大功率可超过1.2W/cm2。用电化学分析方法研究了电堆的静态与动态电性能。发现小于1A/cm2的电流密度有利于电堆活化,而电流密度超过1.5A/cm2会使电堆的稳定性变坏。     

质子交换膜燃料电池膜电极的关键技术

 膜电极是多相物质传输和电化学反应场所,决定着燃料电池的性能、寿命及成本。本文分析膜电极当前技术现状与商业化目标,梳理膜电极分类及经过梯度化膜电极向有序化膜电极发展的技术脉络,介绍近年来超低Pt载量的第三代膜电极-有序化膜电极的新进展,比较各种有序化膜电极制备方法的优缺点。目前有序化膜电极在铂族元素总载量为0.118 mg/cm²下取得的最好性能为861 mW/cm²@0.692 V,0.137 g/kW,成本降至5美元/kW,Q/ΔT值从2013年的1.9下降到1.45。从降低Pt用量及简化燃料电池发电系统、降低系统成本的角度看,自增湿有序化膜电极是未来膜电极开发的重要方向。     

EVOH质子交换膜的制备及表征

以碳酸钾为催化剂由乙烯-乙烯醇共聚物(EVOH)与1,3丙烷磺酸内酯(PS)的磺化反应制得离子聚合物(EVOH-g-SO3K),该离子聚合物透析后通过流延法成膜得到质子交换膜(EVOH-g-SO3H).探讨了离子交换容量(IEC)对质子交换膜传导率的影响,当K2CO3/PS摩尔比为1:2,PS用量为0.03mol,IEC=0.67mmol/g时电导率达到最大值0.003S/cm;研究了S-EVOH与EVOH溶液共混后其质子传导率的变化,当S-EVOH/EVOH的质量比为7∶3时电导率可达到0.012S/cm.研究了质子交换膜的质子传导率,离子交换容量(IEC),热学方面的性能.     

质子交换膜燃料电池启动策略

为了缩短质子交换膜燃料电池启动过程中氢气/空气界面存在的时间并限制电堆启动电压,通过实验研究直接启动、启动前氢气吹扫时间以及启动辅助负载对质子交换膜燃料电池性能影响的差异性,在此基础上提出一种电堆启动时氢气吹扫阳极和启动辅助负载相结合的燃料系统启动控制策略。实验验证了该启动控制策略不仅能限制燃料电池启动时的高电压以及缩短燃料电池启动过程中电堆阳极侧氢气/空气界面的存在时间,还有利于提高单电池的电压均衡性,是一种有效的质子交换膜燃料电池启动控制策略。