Three-dimensional structure prediction of the NAD binding site of proton-pumping transhydrogenase from Escherichia coli

A three-dimensional structure of the NAD site of Escerichia coli transhydrogenase has been predicted. The model is based on analysis of conserved residues among the transhydrogenases from five different sources, homologies with enzymes using NAD as cofactors or substrates, hydrophilicity profiles, and secondary structure predictions. The present model supports the hypothesis that there is one binding site, located relatively close to the N-terminus of the α-subunit. The proposed structure spans residues α145 to α287, and it includes five β-strands and five α-helices oriented in a typical open twisted α/β conformation. The amino acid sequence following the GXGXXG dinucleotide binding consensus sequence (residues α172 to α177) correlates exactly to a typical fingerprint region for ADP binding βαβ folds in dinucleotide binding enzymes. In the model, aspartic acid α195 forms hydrogen bonds to one or both hydroxyl groups on the adenosine ribose sugar moiety. Threonine α196 and alanine α256, located at the end of βB and βD, respectively, create a hydrophobic sandwich with the adenine part of NAD buried inside. The nicotinamide part is located in a hydrophobic cleft between αA and βE. Mutagenesis work has been carried out in order to test the predicted model and to determine whether residues within this domain are important for proton pumping directly. All data support the predicted structure, and no residue crucial for proton pumping Was detected. Since no three-dimensional structure of transhydrogenase has been solved, a well based tertiary structure prediction is of great value for further experimental design in trying to elucidate the mechanism of the energy-linked proton pump. © 1995 Wiley-Liss, Inc.     

The type IV pre-pilin leader peptidase of Xanthomonas campestris pv. campestris is functional without conserved cysteine residues

Type IV pre-pilin leader peptidase was demonstrated to be required for protein secretion, in addition to its involvement in biogenesis of type IV pili. The type IV pre-pilin leader peptidase gene of Xanthomonas campestris pv. campestris was located on a 3 kb Acc l fragment on account of its hybridization with the DNA fragment containing the type IV pre-pilin leader-peptidase gene pilD/xcpA of Pseudomonas aeruginosa . Sequencing of the cloned fragment revealed an open reading frame (ORF) (designated xpsO ) of 287 amino acid residues. A protein with an apparent molecular mass of approximately 32.5 kDa was synthesized in vitro from a DNA fragment containing the xpsO gene. The amino acid sequence shares 50% identity with that of PilD throughout the entire sequence. Among other type IV pre-pilin leader peptidases, XpsO is unique in not having the two conserved -CXXC- motifs in a cytoplasmic domain. Instead, new motifs were noted when the protein was compared with XpsE, which is another member of the extracellular protein-secretion machinery. When the xpsO gene was introduced into the pilD mutant of P. aeruginosa , both the sensitivity against infection with the pilus-specific phage PO4 and the ability to secrete extracellular protein were recovered. Furthermore, immunoblot analysis indicated that the P. aeruginosa pilin was apparently processed in vivo by the xpsO gene product.     

一种免受乙醇干扰的GOD的酶电极

将酶电极应用于发酵糖的测定时。与糖共存的乙醇常常会影响测糖的准确性,通过对葡萄糖氧化酶(GOD)电极的研究,探讨了乙醇对测糖酶电极测定影响,进而研制出抗干扰的GOD酶电极,若是GOD电极的酶膜通过夹心法制备．在乙醇含量为0．1％(V／V)时·即产生显著的影响,使测定结果偏大4．3%,且乙醇的影响随浓度的升高而增大,若用尼龙网固定GOD膜,GOD电极在测定20mmol／L和5mm01／L左右的葡萄糖溶液时,含量高达9％的乙醇仍未对测定产生显著的影响．表现出良好的不受乙醇干扰的特性,并且,该尼龙网GOD电极具有良好的重复性、稳定的响应活性及较长的保存期．     

低高径比喷射环流生化反应器流体力学和发酵性能的研究

对高径比s≤2．5喷射环流生化反应器的流体力学和传质特性进行了系统的研究,选出反应器的最佳结构,关联出氧的体积传递系数(k_La)表达式。在此基础上,进行了谷氨酸发酵试验,摸索出用该设备进行各氨酸发酵的最佳工艺条件,使5批一次性投糖发酵的糖酸转化率达到50％以上。     

背角无齿蚌珍珠囊形成过程的组织学和酶组织化学研究

本文采用Ｈ．Ｅ染色法和酶组织化学方法,对背角无齿蚌珍珠囊形成过程的组织学和酶组织化学的变化情况进行了研究,证实了“初生珍珠囊”的形成和溶解及“次生珍珠囊”的形成,并推没“次生珍珠囊”表皮细胞是由育珠蚌结缔组织转化而来的;观察了育珠手术后九种酶在珍珠囊形成的各个阶段和在珍珠囊和育珠蚌不同部位的变化情况,说明了珍珠囊珙成过程是与复杂的能量代谢,物质代谢及物质转运等有关的生理生化过程。     

拉萨郊区藏族的指纹研究

本文报道了拉萨郊区５１７例（男２２６人,女２９１人）藏族健康人的指纹参数正常值、调查分析了指纹类型、指纹组合、指纹指数和指嵴纹计数。比较了藏族不同居群、不同民族和人种间的差异。结果表明,藏族有自己的指纹特点,又显著蒙古人种的一般特征。     

蛋白质特异性断裂试剂研究进展

蛋白质特异性断裂试剂是近年来发展起来的一些具有特异性断裂肽键功能的化学工具．这些断裂试剂可分为两类,一类通过氧化断裂机制实现蛋白空间结构特异性切割,另一类通过水解断裂机制实现序列特异性切割．蛋白质特异性断裂试剂在蛋白质序列测定,蛋白质的结构与功能研究,蛋白质与核酸相互作用研究以及新型化学治疗药物的合成等方面有着广阔的应用前景．     

新疆10种沙生植物旱生结构的解剖学研究

新疆１０种沙生植物的形态解剖研究表明,它们为适应沙生环境形态结构发生变化。叶器官的形态呈三种类型：叶片退化成膜质或鳞片状,而由同化枝执行光合功能;叶片上下都具栅栏组织,表皮角质膜厚,表皮毛发达,气孔下陷,输导组织和机械组织都发达;叶片肉质化,叶肉组织不分化,贮水组织发达而输导组织不发达。轴器官中厚壁组织发达,围绕维管组织,维管组织内部也有发达的厚壁组织。根中普遍具有周皮,一些植物存在异常的维管组织,部分植物还具有粘液细胞或结晶。沙生植物形成各种旱生结构,以不同的方式适应沙生环境。