Structure of the rotor ring of F-Type Na+-ATPase from Ilyobacter tartaricus

In the crystal structure of the membrane-embedded rotor ring of the sodium ion-translocating adenosine 5'-triphosphate (ATP) synthase of Ilyobacter tartaricus at 2.4 angstrom resolution, 11 c subunits are assembled into an hourglass-shaped cylinder with 11-fold symmetry. Sodium ions are bound in a locked conformation close to the outer surface of the cylinder near the middle of the membrane. The structure supports an ion-translocation mechanism in the intact ATP synthase in which the binding site converts from the locked conformation into one that opens toward subunit a as the rotor ring moves through the subunit a/c interface.     

Mechanical rotation of the c subunit oligomer in ATP synthase (F0F1): direct observation

F0F1, found in mitochondria or bacterial membranes, synthesizes adenosine 5'-triphosphate (ATP) coupling with an electrochemical proton gradient and also reversibly hydrolyzes ATP to form the gradient. An actin filament connected to a c subunit oligomer of F0 was able to rotate by using the energy of ATP hydrolysis. The rotary torque produced by the c subunit oligomer reached about 40 piconewton-nanometers, which is similar to that generated by the gamma subunit in the F1 motor. These results suggest that the gamma and c subunits rotate together during ATP hydrolysis and synthesis. Thus, coupled rotation may be essential for energy coupling between proton transport through F0 and ATP hydrolysis or synthesis in F1.     

High-speed atomic force microscopy reveals rotary catalysis of rotorless F₁-ATPase

F(1) is an adenosine triphosphate (ATP)-driven motor in which three torque-generating β subunits in the α(3)β(3) stator ring sequentially undergo conformational changes upon ATP hydrolysis to rotate the central shaft γ unidirectionally. Although extensive experimental and theoretical work has been done, the structural basis of cooperative torque generation to realize the unidirectional rotation remains elusive. We used high-speed atomic force microscopy to show that the rotorless F(1) still "rotates"; in the isolated α(3)β(3) stator ring, the three β subunits cyclically propagate conformational states in the counterclockwise direction, similar to the rotary shaft rotation in F(1). The structural basis of unidirectionality is programmed in the stator ring. These findings have implications for cooperative interplay between subunits in other hexameric ATPases.     

Oxidative phosphorylation at the fin de siècle

Mitochondria produce most of the energy in animal cells by a process called oxidative phosphorylation. Electrons are passed along a series of respiratory enzyme complexes located in the inner mitochondrial membrane, and the energy released by this electron transfer is used to pump protons across the membrane. The resultant electrochemical gradient enables another complex, adenosine 5'-triphosphate (ATP) synthase, to synthesize the energy carrier ATP. Important new mechanistic insights into oxidative phosphorylation have emerged from recent three-dimensional structural analyses of ATP synthase and two of the respiratory enzyme complexes, cytochrome bc1 and cytochrome c oxidase. This work, and new enzymological studies of ATP synthase's unusual catalytic mechanism, are reviewed here.     

The E. coli BtuCD structure: a framework for ABC transporter architecture and mechanism

The ABC transporters are ubiquitous membrane proteins that couple adenosine triphosphate (ATP) hydrolysis to the translocation of diverse substrates across cell membranes. Clinically relevant examples are associated with cystic fibrosis and with multidrug resistance of pathogenic bacteria and cancer cells. Here, we report the crystal structure at 3.2 angstrom resolution of the Escherichia coli BtuCD protein, an ABC transporter mediating vitamin B12 uptake. The two ATP-binding cassettes (BtuD) are in close contact with each other, as are the two membrane-spanning subunits (BtuC); this arrangement is distinct from that observed for the E. coli lipid flippase MsbA. The BtuC subunits provide 20 transmembrane helices grouped around a translocation pathway that is closed to the cytoplasm by a gate region whereas the dimer arrangement of the BtuD subunits resembles the ATP-bound form of the Rad50 DNA repair enzyme. A prominent cytoplasmic loop of BtuC forms the contact region with the ATP-binding cassette and appears to represent a conserved motif among the ABC transporters.     

玉米秸秆接触物理参数测定与离散元仿真标定

为提高离散元方法模拟玉米秸秆揉碎过程的准确性,试验测定玉米秸秆与揉碎机锤片、玉米秸秆与玉米秸秆的接触参数。以径向堆积角相对误差值为评价指标,应用正交方法标定玉米秸秆离散元仿真需要输入的接触参数。结果表明:玉米秸秆与揉碎机锤片碰撞恢复系数、静摩擦因数、滚动摩擦因数分别设置为0.663、0.226、0.119;玉米秸秆与玉米秸秆碰撞恢复系数、静摩擦因数、滚动摩擦因数分别设置为0.485、0.142、0.078,此参数组合为玉米秸秆离散元仿真接触参数的最优方案,仿真结果与试验结果的相对误差为8.127%。     

荻草茎秆动态润湿性能及表面自由能

荻草Miscanthus sacchariflorus是一种高大直立的多年生高生物量草类。通过对荻草茎秆动态润湿模型、表面自由能的研究,探索荻草茎秆被胶黏剂胶合及作为人造板原材料的可能性。运用接触角测试仪,分别测定脲醛树脂(UF),三聚氰胺改性脲醛树脂(MUF)和酚醛树脂(PF)在荻草茎秆内、外表面的接触角,拟合出动态润湿模型。利用扩散渗透系数K,比较3种胶黏剂对荻草茎秆的润湿能力。运用表面张力仪测试荻草茎秆内、外表面自由能。结果表明:3种胶黏剂在荻草内、外表面的润湿模型,能很好地模拟接触角随时间变化的关系;3种胶黏剂在荻草茎秆表面的润湿性能为:三聚氰胺改性脲醛树脂<脲醛树脂<酚醛树脂;荻草茎秆的平均自由能约为47.49 mJ·m-2,与木材相近,高于麦秸。     

Crystal structures of the adenylate sensor from fission yeast AMP-activated protein kinase

The 5'-AMP (adenosine monophosphate)-activated protein kinase (AMPK) coordinates metabolic function with energy availability by responding to changes in intracellular ATP (adenosine triphosphate) and AMP concentrations. Here, we report crystal structures at 2.9 and 2.6 A resolution for ATP- and AMP-bound forms of a core alphabetagamma adenylate-binding domain from the fission yeast AMPK homolog. ATP and AMP bind competitively to a single site in the gamma subunit, with their respective phosphate groups positioned near function-impairing mutants. Unexpectedly, ATP binds without counterions, amplifying its electrostatic effects on a critical regulatory region where all three subunits converge.     

玉米幼根接种禾谷镰刀菌后的转录组分析

为研究禾谷镰刀菌侵染后玉米幼根内转录组的变化情况,利用RNAseq对禾谷镰刀菌接种后6和18h的感病玉米自交系Y331分别进行转录组测序,分析接种后玉米幼根内差异表达的基因。结果表明:接种禾谷镰刀菌后,玉米幼根内共有5 153个基因差异表达;在接种后上调表达的聚类中,多种抗病过程相关的基因显著富集,包括苯丙烷类次生代谢物质合成过程的基因以及植物激素水杨酸(Salicylic acid,SA)、茉莉酸(Jasmonic acid,JA)和乙烯(Ethylene,ET)合成、响应及信号介导途径的基因;在下调表达的聚类中,植物生长发育、基础物质和能量代谢相关过程的基因显著富集。进一步分析发现:ET相关过程的基因在聚类1和聚类6中显著富集,均在接种后6h前上调表达;苯丙烷类次生代谢物合成途径相关基因和镰刀烯醇毒素(Deoxynivalenol,DON)解毒相关基因在接种后上调表达。研究表明,SA、JA/ET、苯丙烷类次生代谢物和DON解毒基因在玉米和禾谷镰刀菌互作中发挥重要作用。     

玉米苯丙烷类次生代谢物与玉米对茎腐病抗性的关系

对抗病玉米品种陕单931和感病玉米品种西农11号在抽雄初期接种禾谷镰刀菌孢子悬浮液,于接种后测定茎秆髓部组织内苯丙烷类次生代谢物木质素和绿原酸的变化。结果表明,玉米植株原生木质素与玉米对茎腐病的抗性无关,仅诱导产生的木质素在玉米的抗病性中起作用。并证实玉米植株受到镰刀菌侵染后可产生对镰刀菌有抑制作用的物质。     

Structure of the rotor of the V-Type Na+-ATPase from Enterococcus hirae

The membrane rotor ring from the vacuolar-type (V-type) sodium ion-pumping adenosine triphosphatase (Na+-ATPase) from Enterococcus hirae consists of 10 NtpK subunits, which are homologs of the 16-kilodalton and 8-kilodalton proteolipids found in other V-ATPases and in F1Fo- or F-ATPases, respectively. Each NtpK subunit has four transmembrane alpha helices, with a sodium ion bound between helices 2 and 4 at a site buried deeply in the membrane that includes the essential residue glutamate-139. This site is probably connected to the membrane surface by two half-channels in subunit NtpI, against which the ring rotates. Symmetry mismatch between the rotor and catalytic domains appears to be an intrinsic feature of both V- and F-ATPases.     

降解棉秆纤维素分解菌的筛选及其降解特性研究

从土壤中筛选到两株能降解棉秆的纤维素分解菌M59、F115,通过构建酶活曲线确定了发酵周期,并进一步检测了不同氮源、不同浓度对菌株降解棉秆酶活的影响。结果表明,M59和F115均能有效降解纤维素,在刚果红纤维素平板上形成的水解圈直径〉0.5cm,降解滤纸的CMC酶活〉7.8 U.ml-1,FPA酶活〉3.9 U.ml-1;两菌株均能降解棉秆且维持较高的酶活（CMC酶活〉7.0 U.ml-1,FPA酶活〉2.5 U.ml-1）,发酵周期均确定为10d;氮源及其浓度对两菌株降解棉秆的CMC酶活和FPA酶活影响显著,以0.5%的酵母粉、蛋白胨作氮源,菌株M59的CMC酶活和FPA酶活、F115的CMC酶活均最高,可确定0.5%酵母粉、蛋白胨为两株纤维素分解菌较适宜的氮源。     

F_1菌株对玉米秸秆木质素和纤维素降解能力的研究

为了探讨F1菌株的产酶活性及其对玉米秸秆中木质素和纤维素的降解能力,以羧甲基纤维素钠为诱导物配制培养基Ⅰ,以不加羧甲基纤维素钠为对照配制培养基Ⅱ,分别接种F1菌株,在25℃下培养诱导产生纤维素降解酶Cx,用分光光度法测定不同时间所产生的酶活。将培养5 d的F1菌株以10%的接种量转接到玉米秸秆固体发酵培养基上培养,在5 d1、0 d、15 d2、0 d用差重法测定F1菌株对玉米秸秆中半纤维素、纤维素和木质素的降解能力。结果表明,以羧甲基纤维素钠为诱导物培养基,F1菌株能够产生Cx酶,而且在第5天时酶活性最强。F1菌株在玉米秸秆发酵培养基中的生长情况良好,培养前15 d,F1菌株对纤维素和木质素的降解都很快,之后对纤维素的降解速率明显降低甚至停止降解。培养20 d,对半纤维素、纤维素和木质素的累计降解速率为3.5%、10.5%、7.0%。     

西葫芦果柄长主基因-多基因混合遗传分析

[目的]为西葫芦果柄长育种提供依据。[方法]选用西葫芦自交系配制q-1×23-4G（组合1）和q-1×A-7（组合2）2个组合,构建P1、F1、P2、BC1、BC2和F2 6个家系世代群体,应用植物数量性状主基因-多基因混合遗传模型对该6个世代群体果柄长进行多世代联合分析。[结果]2个组合西葫芦的果柄长性状遗传为加性-显性-上位性2对主基因（B-1）遗传模型;2个组合F2的主基因遗传率较低,环境影响相对较大。[结论]西葫芦果柄长育种宜采用个体选择法,可在晚世代选择。     

基于Plackett-Burman试验设计与响应面法优化玉米秸秆离散元模型

为了减小免耕播种机防堵装置数值模拟的误差,结合物理试验与仿真试验进行玉米秸秆离散元参数的标定。基于Hertz-Mindlin with bonding 接触模型建立了玉米秸秆离散元模型,以物理试验与仿真试验中临界载荷的相对误差为试验指标进行仿真单轴压缩试验;采用Plackett-Burman试验筛选出具有显著性影响的试验因素;采用Box-Behnken响应面法对玉米秸秆离散元模型进行优化。试验结果表明:接触半径和单位面积切向刚度对试验指标影响极显著(P<0.01),单位面积法向刚度对试验指标影响显著(0.01<P<0.05);玉米秸秆的最优离散元参数为,接触半径1.2 mm、单位面积法向刚度9.361×10⁷ N/m³、单位面积切向刚度9.845×10⁷ N/m³,在此条件下,仿真压缩试验的临界载荷为950.2 N,与物理试验值935.4 N的相对误差为1.58%,验证了参数的可靠性。标定的玉米秸秆离散元模型可用于免耕播种机防堵装置的数值模拟,为防堵装置结构优化改进提供了依据。     

玉米植株抗倒伏性状的遗传分析

以2个茎秆强度不同的自交系82 -3和W1为基础材料,通过杂交与自交,对双亲、F1及F2等不同群体株高、穗位高、茎秆强度等植株性状进行田间调查和遗传分析.结果表明,双亲及其杂交后代各性状差异明显,与W1相比,亲本82 -3表现株矮、穗位低,但根系差,茎秆强度也低;杂种F1除穗位节的节间强度表现负向杂种优势外,其它性状均...     

Observation of a membrane fusion intermediate structure

We report the observation of a phase of phospholipid that contains a structure similar to the commonly postulated interbilayer state that is crucial to membrane fusion. The widely accepted model for membrane fusion suggests that there is an intermediate state in which the two contacting monolayers become continuous via an hourglass-shaped structure called a stalk. Many efforts have been made to estimate the free energy for such a state in order to understand the functionality of membrane fusion proteins and to define key parameters in energy estimates. The observation of the stalk structure supports the stalk hypothesis for membrane fusion and enables the measurement of these parameters experimentally.     

玉米穗,茎腐病病原学相互关系及发病条件的研究

玉米穗、茎腐病病原学的相互关系研究表明：其优势致病菌禾谷镰孢菌和串珠镰抱菌均可交互侵染引起这两种病害,并且能从植株的外侧和内部由基部向穗部传染,初步证实引起穗腐病的镰孢菌是来自茎腐病的镰孢菌。穗、茎腐病的发病条件试验结果显示,玉米茎腐病与玉米品种的抗性、生育期、种植密度、地势、降水和施肥等的关系极为密切,而玉米穗腐病与玉米品种的抗性、土壤类型、种植形式和螟害的关系密切。通过施用钾肥和防治玉米螟可有效地减轻玉米茎腐病和穗腐病的危害。     

PKA type IIalpha holoenzyme reveals a combinatorial strategy for isoform diversity

The catalytic (C) subunit of cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) is inhibited by two classes of regulatory subunits, RI and RII. The RII subunits are substrates as well as inhibitors and do not require adenosine triphosphate (ATP) to form holoenzyme, which distinguishes them from RI subunits. To understand the molecular basis for isoform diversity, we solved the crystal structure of an RIIalpha holoenzyme and compared it to the RIalpha holoenzyme. Unphosphorylated RIIalpha(90-400), a deletion mutant, undergoes major conformational changes as both of the cAMP-binding domains wrap around the C subunit's large lobe. The hallmark of this conformational reorganization is the helix switch in domain A. The C subunit is in an open conformation, and its carboxyl-terminal tail is disordered. This structure demonstrates the conserved and isoform-specific features of RI and RII and the importance of ATP, and also provides a new paradigm for designing isoform-specific activators or antagonists for PKA.     

玉米穗,茎腐病禾谷镰孢菌和串珠镰孢菌同工酶分析

对沈阳、河北、吉林、北京等地区玉米穗、茎腐病禾谷镰孢菌和串珠镰孢菌进行了可溶性蛋白和同工酶聚丙烯酰胺凝胶电泳,以研究玉米穗、茎腐病同种镰孢菌是否存在寄生专化性或生理分化现象。结果表明：各地区的玉米穗、茎腐病串珠镰孢菌同源性高,亲缘关系近,相似性系数在0．86～0．98之间,菌株间差异仅表现为个体间多样性。而两种病害禾谷镰孢菌亲缘关系困地区不同而有差异,河北、吉林两地区穗、茎腐禾谷镰孢菌间均具有很近的亲缘关系,相似性系数在0．93以上;但沈阳地区禾谷镰孢菌亲缘关系相对较远,相似性系数仅为0．65～0．67。