Organization of iron-sulfur clusters in respiratory complex I

Complex I of respiratory chains plays a central role in bioenergetics and is implicated in many human neurodegenerative diseases. An understanding of its mechanism requires a knowledge of the organization of redox centers. The arrangement of iron-sulfur clusters in the hydrophilic domain of complex I from Thermus thermophilus has been determined with the use of x-ray crystallography. One binuclear and six tetranuclear clusters are arranged, maximally 14 angstroms apart, in an 84-angstrom-long electron transfer chain. The binuclear cluster N1a and the tetranuclear cluster N7 are not in this pathway. Cluster N1a may play a role in the prevention of oxidative damage. The structure provides a framework for the interpretation of the large amounts of data accumulated on complex I.     

Structure of hexameric DnaB helicase and its complex with a domain of DnaG primase

The complex between the DnaB helicase and the DnaG primase unwinds duplex DNA at the eubacterial replication fork and synthesizes the Okazaki RNA primers. The crystal structures of hexameric DnaB and its complex with the helicase binding domain (HBD) of DnaG reveal that within the hexamer the two domains of DnaB pack with strikingly different symmetries to form a distinct two-layered ring structure. Each of three bound HBDs stabilizes the DnaB hexamer in a conformation that may increase its processivity. Three positive, conserved electrostatic patches on the N-terminal domain of DnaB may also serve as a binding site for DNA and thereby guide the DNA to a DnaG active site.     

嗜热链球菌胞外多糖纯化条件的研究

[目的]探索一种分离纯化嗜热链球菌胞外多糖的途径。[方法]采用嗜热链球菌为材料,以MRS培养基为基础培养基,生产嗜热链球菌胞外多糖。采用酶法、TCA法、Sevag法、酶＋TCA法、酶＋Sevag法、TCA＋Sevag法、酶＋TCA＋Sevag法研究对嗜热链球菌胞外多糖去蛋白的影响,对得到的粗多糖用DEAE-Cellulose离子交换柱和葡聚糖G-100凝胶柱进行了分离,并用紫外光谱扫描对其的纯度进行鉴定。[结果]粗多糖用蒸馏水和0.1 mol/L NaC l通过DEAE-Cellulose离子交换柱可以分为中性多糖和酸性多糖,用葡聚糖G-100凝胶柱对酸性多糖的纯度进行了鉴定,得到单一的峰,紫外光谱扫描对酸性多糖进行全波长扫描,酸性多糖中不含蛋白质和核酸,证明得到的酸性多糖为纯多糖。[结论]酶＋Sevag法为最佳去蛋白法。     

猪呼吸道疾病综合征的综合防制

猪呼吸道疾病综合征(Porcine respiratory dis-ease complex,PRDC)是由多种病因(病毒、细菌、寄生虫、中毒、环境应激和猪体免疫力低下等)引起的猪呼吸障碍性疾病的总称。近年来,猪呼吸道疾病的发生和流行日益严重,主要多见于断奶仔猪、哺乳仔猪及刚进入育成舍的猪只。该病无明显的季节性,尤以冬、春季节多发,发病率可达30%~50%,死亡率高达100%;其他季节发病率为10%左右。1发病的主要因素PRDC涉及多种病原体,包括原发性感染疾病和继发性感染疾病。原发性感染疾病包括猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒等病毒性疾病以及猪喘气病、…     

Structure of the S15,S6,S18-rRNA complex: assembly of the 30S ribosome central domain

The crystal structure of a 70-kilodalton ribonucleoprotein complex from the central domain of the Thermus thermophilus 30S ribosomal subunit was solved at 2.6 angstrom resolution. The complex consists of a 104-nucleotide RNA fragment composed of two three-helix junctions that lie at the end of a central helix, and the ribosomal proteins S15, S6, and S18. S15 binds the ribosomal RNA early in the assembly of the 30S ribosomal subunit, stabilizing a conformational reorganization of the two three-helix junctions that creates the RNA fold necessary for subsequent binding of S6 and S18. The structure of the complex demonstrates the central role of S15-induced reorganization of central domain RNA for the subsequent steps of ribosome assembly.     

Structure of the LDL receptor extracellular domain at endosomal pH

The low-density lipoprotein receptor mediates cholesterol homeostasis through endocytosis of lipoproteins. It discharges its ligand in the endosome at pH < 6. In the crystal structure at pH = 5.3, the ligand-binding domain (modules R2 to R7) folds back as an arc over the epidermal growth factor precursor homology domain (the modules A, B, beta propeller, and C). The modules R4 and R5, which are critical for lipoprotein binding, associate with the beta propeller via their calcium-binding loop. We propose a mechanism for lipoprotein release in the endosome whereby the beta propeller functions as an alternate substrate for the ligand-binding domain, binding in a calcium-dependent way and promoting lipoprotein release.     

Structure of the RNA polymerase domain of E. coli primase

All cellular organisms use specialized RNA polymerases called "primases" to synthesize RNA primers for the initiation of DNA replication. The high-resolution crystal structure of a primase, comprising the catalytic core of the Escherichia coli DnaG protein, was determined. The core structure contains an active-site architecture that is unrelated to other DNA or RNA polymerase palm folds, but is instead related to the "toprim" fold. On the basis of the structure, it is likely that DnaG binds nucleic acid in a groove clustered with invariant residues and that DnaG is positioned within the replisome to accept single-stranded DNA directly from the replicative helicase.     

Structure of an oxygen-carrying cobalt complex

Bis(3-fluorosalicylaldehyde) ethylenediimine cobalt(II), a reversible oxygen-carrying compound, has been crystallized as a tetramer with two oxygen molecules bound to the four cobalt atomns, each oxygen molecule bridging between two metal atoms. These dimers are further linked by two bonds between the oxygen atom of the salicylaldehyde and the cobalt atom. The oxygen-to-oxygen distance, 1.21 angstroms in molecular oxygen, has increased to 1.308(28) angstroms (where the number in parentheses is the estimated standard deviation) in this compound, the shortest distance yet observed in sutch a bridging arrangement.     

水生栖热菌FL-03海藻糖合酶的分离纯化及特性研究

将从水生栖热菌FL-03中获得的海藻糖合酶粗酶液经硫酸铵分级沉淀、DEAE Sepharose CL-6B离子交换层析S、ephacryl S-200HR凝胶过滤层析后,纯化68.48倍,产率为18.4%。研究表明:该酶分子量约为101 kD,最适反应温度为60℃,最适pH为7.0,在40~80℃、pH 6.0~9.0范围内具有较高的反应活性和稳定性。Fe3 对该酶具有一定的激活作用,Cu2 、Tris、Zn2 对该酶具有较强的抑制作用。该酶具有高度的底物特异性,只能专一地将麦芽糖转化为海藻糖。在40℃4、8 h条件下,麦芽糖转化为海藻糖的转化率最大可达到79%。     

牛呼吸疾病综合征诊断方法的研究进展

牛呼吸疾病综合征是危害国内外养牛业的一种重要传染病,致病因素包括病毒、细菌以及支原体等病原体和多种应激。养殖一线人员常常只能通过观察牛的呼吸异常而做出初步诊断,对病原体诊断的准确率低。而且,由于该病具有多病因混合感染的特征,实验室诊断也常常针对性不强,导致难以对患病动物的治疗效果及预后做出精准判断。为了给牛呼吸疾病综合征的早期诊断、及时治疗和综合防控提供参考依据,本文就牛呼吸疾病综合征的病原学、流行和危害以及诊断方法进行综述,包括常见病原,临床诊断、分子生物学诊断等常用诊断方法,以及含有急性期蛋白和应激相关激素的宿主生物标志物诊断、转录组学诊断等新型诊断方法,简述其实际应用情况和优缺点,并对未来诊断方法的方向及待解决问题等提出展望。     

Structure of SARS coronavirus spike receptor-binding domain complexed with receptor

The spike protein (S) of SARS coronavirus (SARS-CoV) attaches the virus to its cellular receptor, angiotensin-converting enzyme 2 (ACE2). A defined receptor-binding domain (RBD) on S mediates this interaction. The crystal structure at 2.9 angstrom resolution of the RBD bound with the peptidase domain of human ACE2 shows that the RBD presents a gently concave surface, which cradles the N-terminal lobe of the peptidase. The atomic details at the interface between the two proteins clarify the importance of residue changes that facilitate efficient cross-species infection and human-to-human transmission. The structure of the RBD suggests ways to make truncated disulfide-stabilized RBD variants for use in the design of coronavirus vaccines.     

水葫芦水提取物的抗氧化活性研究

采用热水浸提新鲜水葫芦得到水葫芦粗提液,对粗提液的还原力以及DPPH自由基、羟自由基、超氧阴离子自由基的清除能力和抗脂质过氧化能力进行测定。结果表明,当水葫芦粗提液的蛋白与Vc浓度相同时,水葫芦粗提液的抗氧化活性等于或高于Vc,其清除羟自由基和还原力远高于Vc,且抗氧化活性随提取液浓度的升高而增大,说明水葫芦水粗提物具有一定的抗氧化活性。     

酸奶中嗜热乳酸链球菌的分离鉴定及益生特性研究

[目的]探讨酸奶中嗜热乳酸链球菌的益生特性。[方法]利用MRS琼脂培养基从酸奶中分离嗜热乳酸链球菌,通过糖发酵试验对其进行生化鉴定。研究嗜热乳酸链球菌的抑菌特性和胆盐耐受性,并分析不同pH值对嗜热乳酸链球菌的的影响。[结果]镜检结果表明该菌为革兰氏阳性,经生化鉴定可确定该菌为嗜热乳酸链球菌。嗜热乳酸链球菌对人体肠道内的常见病原菌有抑制作用,尤其是对大肠杆菌和金黄色葡萄球菌的抑制作用明显。嗜热乳酸链球菌可以耐受人体肠道的胆盐环境,并在其特定部位生存。嗜热乳酸链球菌可耐受强酸,能在胃酸中存活,在近中性pH值环境中生长较好。[结论]嗜热乳酸链球菌可在人体肠道内发挥益生作用,保护机体免受病原菌的侵害。     

Unexpected modes of PDZ domain scaffolding revealed by structure of nNOS-syntrophin complex

The PDZ protein interaction domain of neuronal nitric oxide synthase (nNOS) can heterodimerize with the PDZ domains of postsynaptic density protein 95 and syntrophin through interactions that are not mediated by recognition of a typical carboxyl-terminal motif. The nNOS-syntrophin PDZ complex structure revealed that the domains interact in an unusual linear head-to-tail arrangement. The nNOS PDZ domain has two opposite interaction surfaces-one face has the canonical peptide binding groove, whereas the other has a beta-hairpin "finger." This nNOS beta finger docks in the syntrophin peptide binding groove, mimicking a peptide ligand, except that a sharp beta turn replaces the normally required carboxyl terminus. This structure explains how PDZ domains can participate in diverse interaction modes to assemble protein networks.     

Structure of a NHEJ polymerase-mediated DNA synaptic complex

Nonhomologous end joining (NHEJ) is a critical DNA double-strand break (DSB) repair pathway required to maintain genome stability. Many prokaryotes possess a minimalist NHEJ apparatus required to repair DSBs during stationary phase, composed of two conserved core proteins, Ku and ligase D (LigD). The crystal structure of Mycobacterium tuberculosis polymerase domain of LigD mediating the synapsis of two noncomplementary DNA ends revealed a variety of interactions, including microhomology base pairing, mismatched and flipped-out bases, and 3' termini forming hairpin-like ends. Biochemical and biophysical studies confirmed that polymerase-induced end synapsis also occurs in solution. We propose that this DNA synaptic structure reflects an intermediate bridging stage of the NHEJ process, before end processing and ligation, with both the polymerase and the DNA sequence playing pivotal roles in determining the sequential order of synapsis and remodeling before end joining.     

Structure of the protease domain of memapsin 2 (beta-secretase) complexed with inhibitor

Memapsin 2 (beta-secretase) is a membrane-associated aspartic protease involved in the production of beta-amyloid peptide in Alzheimer's disease and is a major target for drug design. We determined the crystal structure of the protease domain of human memapsin 2 complexed to an eight-residue inhibitor at 1.9 angstrom resolution. The active site of memapsin 2 is more open and less hydrophobic than that of other human aspartic proteases. The subsite locations from S4 to S2' are well defined. A kink of the inhibitor chain at P2' and the change of chain direction of P3' and P4' may be mimicked to provide inhibitor selectivity.     

Structure of the VHL-ElonginC-ElonginB complex: implications for VHL tumor suppressor function

Mutation of the VHL tumor suppressor is associated with the inherited von Hippel-Lindau (VHL) cancer syndrome and the majority of kidney cancers. VHL binds the ElonginC-ElonginB complex and regulates levels of hypoxia-inducible proteins. The structure of the ternary complex at 2.7 angstrom resolution shows two interfaces, one between VHL and ElonginC and another between ElonginC and ElonginB. Tumorigenic mutations frequently occur in a 35-residue domain of VHL responsible for ElonginC binding. A mutational patch on a separate domain of VHL indicates a second macromolecular binding site. The structure extends the similarities to the SCF (Skp1-Cul1-F-box protein) complex that targets proteins for degradation, supporting the hypothesis that VHL may function in an analogous pathway.     

Structure of the DNA-Eco RI endonuclease recognition complex at 3 A resolution

The crystal structure of the complex between Eco RI endonuclease and the cognate oligonucleotide TCGCGAATTCGCG provides a detailed example of the structural basis of sequence-specific DNA-protein interactions. The structure was determined, to 3 A resolution, by the ISIR (iterative single isomorphous replacement) method with a platinum isomorphous derivative. The complex has twofold symmetry. Each subunit of the endonuclease is organized into an alpha/beta domain consisting a five-stranded beta sheet, alpha helices, and an extension, called the "arm," which wraps around the DNA. The large beta sheet consists of antiparallel and parallel motifs that form the foundations for the loops and alpha helices responsible for DNA strand scission and sequence-specific recognition, respectively. The DNA cleavage site is located in a cleft that binds the DNA backbone in the vicinity of the scissile bond. Sequence specificity is mediated by 12 hydrogen bonds originating from alpha helical recognition modules. Arg200 forms two hydrogen bonds with guanine while Glu144 and Arg145 form four hydrogen bonds to adjacent adenine residues. These interactions discriminate the Eco RI hexanucleotide GAATTC from all other hexanucleotides because any base substitution would require rupture of at least one of these hydrogen bonds.     

Crystal Structure of the Solid Electrolyte, RbAg4I5

The crystal structure of the solid electrolyte RbAg(4)I(5) has been determined from single-crystal x-ray diffraction counter data. There are four RbAg(4)I(5) in a cubic unit cell with a = 11.24 A. The structure refinement, by least squares, is based on space group P4(1)3 (O(7)). The arrangement of the iodide ions is similar to that of the manganese atoms in beta-manganese, and provides 56 iodide tetrahedra per unit cell, which share faces in such manner as to provide diffusion paths for the silver ions. The occurrence of the two low-temperature phases of RbAg(4)I(5) has been established by x-ray diffraction and optical examination.