Structure of the membrane protein FhaC: a member of the Omp85-TpsB transporter superfamily

In Gram-negative bacteria and eukaryotic organelles, beta-barrel proteins of the outer membrane protein 85-two-partner secretion B (Omp85-TpsB) superfamily are essential components of protein transport machineries. The TpsB transporter FhaC mediates the secretion of Bordetella pertussis filamentous hemagglutinin (FHA). We report the 3.15 A crystal structure of FhaC. The transporter comprises a 16-stranded beta barrel that is occluded by an N-terminal alpha helix and an extracellular loop and a periplasmic module composed of two aligned polypeptide-transport-associated (POTRA) domains. Functional data reveal that FHA binds to the POTRA 1 domain via its N-terminal domain and likely translocates the adhesin-repeated motifs in an extended hairpin conformation, with folding occurring at the cell surface. General features of the mechanism obtained here are likely to apply throughout the superfamily.     

Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface

Foreign DNA fragments can be inserted into filamentous phage gene III to create a fusion protein with the foreign sequence in the middle. The fusion protein is incorporated into the virion, which retains infectivity and displays the foreign amino acids in immunologically accessible form. These "fusion phage" can be enriched more than 1000-fold over ordinary phage by affinity for antibody directed against the foreign sequence. Fusion phage may provide a simple way of cloning a gene when an antibody against the product of that gene is available.     

M13噬菌体衣壳蛋白的研究应用

M13噬菌体是一种丝状噬菌体,内有一个环状单链DNA分子,它由约2 800个PⅧ蛋白组成的圆柱形衣壳包围环状单链DNA分子,其中5个PⅦ和5个PⅪ蛋白在噬菌体的一端,另一端有5个PⅥ蛋白和5个PⅢ蛋白。这些衣壳蛋白参与着噬菌体的感染、组装等功能。文中对这5种衣壳蛋白在噬菌体展示技术的研究应用进行了综述。     

噬菌体抗体库技术在水产养殖中的研究进展

噬菌体抗体库技术是将目的基因克隆到噬菌体的外壳蛋白基因组中,转染宿主菌后,使目的基因编码蛋白和外壳蛋白以融合蛋白的形式表达,呈现在噬菌体表面,通过一系列的筛选获得特异性噬菌体抗体分子.介绍了噬菌体抗体库技术在水产养殖过程中的疾病防控、水环境监测、水产品安全控制以及与抗体芯片结合研究等方面的研究进展.     

Root Hairs, Cuticle, and Pits

The filamentous roots of mustard (Raphanus sativus), radish (Brassica nigra), squash (Cucurbita pepo), and wheat (Triticum aestivum) are covered throughout their length with living nucleated root hairs which may measure 1600 micro or more. The outer walls of piliferous and nonpiliferous cells consist of successive layers of mucilage, cutin, and the cellulose-pectic framework of the cell. Plasmodesmata and pits occur on all cell walls. Under the electron microscope individual pores and pits in the microfibrillar wall are evident throughout the length of the root hair. The "semipermeable membrane" of the root hair zone is thus structurally complex.     

Role of a highly conserved bacterial protein in outer membrane protein assembly

After transport across the cytoplasmic membrane, bacterial outer membrane proteins are assembled into the outer membrane. Meningococcal Omp85 is a highly conserved protein in Gram-negative bacteria, and its homolog Toc75 is a component of the chloroplast protein-import machinery. Omp85 appeared to be essential for viability, and unassembled forms of various outer membrane proteins accumulated upon Omp85 depletion. Immunofluorescence microscopy revealed decreased surface exposure of outer membrane proteins, which was particularly apparent at the cell-division planes. Thus, Omp85 is likely to play a role in outer membrane protein assembly.     

Changes in the cell membranes of the bullfrog gastric mucosa with Acid secretion

The effective area, resistance, and configuration of the apical and basolateral cell membranes of the bullfrog gastric mucosa were studied as a function of acid secretion rate, by alternating-current impedance methods. The drop in transepithelial resistance with acid secretion is attributed to the great increase in apical membrane area (hence conductance) associated with tubulovesicles. There is no evidence of a change in basolateral membrane resistance or of apical membrane premeability per unit area.     

Negative Effects of Sludge Bulking in Membrane Bio-Reactor

The membrane bio-reactor (MBR)is a newbioche- mical reaction system. The study on its applications in treatingvarious types ofwaste streams, such as domestic wastewater, industrial wastewater and human excrement has therefore attracted great attention[1-3…     

Outer membrane active transport: structure of the BtuB:TonB complex

In Gram-negative bacteria, the import of essential micronutrients across the outer membrane requires a transporter, an electrochemical gradient of protons across the inner membrane, and an inner membrane protein complex (ExbB, ExbD, TonB) that couples the proton-motive force to the outer membrane transporter. The inner membrane protein TonB binds directly to a conserved region, called the Ton-box, of the transporter. We solved the structure of the cobalamin transporter BtuB in complex with the C-terminal domain of TonB. In contrast to its conformations in the absence of TonB, the Ton-box forms a beta strand that is recruited to the existing beta sheet of TonB, which is consistent with a mechanical pulling model of transport.     

C-terminal signal sequence promotes virulence factor secretion in Mycobacterium tuberculosis

Mycobacterium tuberculosis uses the ESX-1/Snm system [early secreted antigen 6 kilodaltons (ESAT-6) system 1/secretion in mycobacteria] to deliver virulence factors into host macrophages during infection. Despite its essential role in virulence, the mechanism of ESX-1 secretion is unclear. We found that the unstructured C terminus of the CFP-10 substrate was recognized by Rv3871, a cytosolic component of the ESX-1 system that itself interacts with the membrane protein Rv3870. Point mutations in the signal that abolished binding of CFP-10 to Rv3871 prevented secretion of the CFP-10 (culture filtrate protein, 10 kilodaltons)/ESAT-6 virulence factor complex. Attachment of the signal to yeast ubiquitin was sufficient for secretion from M. tuberculosis cells, demonstrating that this ESX-1 signal is portable.     

Voltage- and tension-dependent lipid mobility in the outer hair cell plasma membrane

The mechanism responsible for electromotility of outer hair cells in the ear is unknown but is thought to reside within the plasma membrane. Lipid lateral diffusion in the outer hair cell plasma membrane is a sigmoidal function of transmembrane potential and bathing media osmolality. Cell depolarization or hyposmotic challenge shorten the cell and reduce membrane fluidity by half. Changing the membrane tension with amphipathic drugs results in similar reductions. These dynamic changes in membrane fluidity represent the modulation of membrane tension by lipid-protein interactions. The voltage dependence may be associated with the force-generating motors that contribute to the exquisite sensitivity of mammalian hearing.     

Mediation of the attachment or fusion step in vesicular transport by the GTP-binding Ypt1 protein

The function of the guanosine triphosphate (GTP)-binding protein Ypt1 in regulating vesicular traffic was studied in a cell-free system that reconstitutes transport from the endoplasmic reticulum to the Golgi. Blocking the Ypt1 protein activity resulted in accumulation of vesicles that act as an intermediate passing between the two compartments. The Ypt1 protein was found on the outer side of these vesicles. The transport process is completed by fusion of these vesicles with the acceptor compartment, and Ypt1 protein activity was needed for this step. Thus, a specific GTP-binding protein is required for either attachment or fusion (or both) of secretory vesicles with the acceptor compartment during protein secretion.     

苎麻韧皮纤维超微结构的观察

[目的]研究苎麻(Boehmeria nivea L.Guad)韧皮纤维的超微结构。[方法]对苎麻茎韧皮纤维进行光学显微镜和透射电子显微镜观察;并对苎麻单纤维进行组织离解,用光学显微镜进行观察。茎韧皮纤维显微观察结果表明:苎麻韧皮纤维细胞壁横切面分为胞间层、初生壁和次生壁3个层次和细胞腔;次生壁大多为1层,即次生壁外层(S1),并且具有许多同心层次;少数纤维具有2～3层,即次生壁外层、中层(S2)和内层(S3);茎基部纤维次生壁多为2～3层,中部纤维次生壁多为1层,苎麻纤维次生壁层次存在位置差异;苎麻纤维是由若干个纤维细胞相互连接而成,纤维细胞之间部分纤维细胞端壁溶解,部分具细胞膜和初生壁,部分具次生壁。苎麻单纤维光学显微镜结果表明:苎麻单纤维细丝状,纤维细胞壁(次生壁)由Z型微纤丝和S型微纤丝组成,具节和节间,少数纤维具有明显的节状加厚现象;中央略大,两端尖削,或钝圆形、二叉状、火炬状等各种形状。[结论]该研究可为生产高品质的苎麻产品奠定基础。     

Proton motive force involved in protein transport across the outer membrane of Aeromonas salmonicida

Many Gram-negative bacteria export proteins to the exterior. Some of these proteins are first secreted into the periplasm and then cross the outer membrane in a separate step. The source of energy required for the translocation is unknown. Export of the extracellular protein proaerolysin from the periplasm through the outer membrane of Aeromonas salmonicida is inhibited by a proton ionophore and by low extracellular pH. One possible explanation of these results is that a proton gradient across the outer membrane is required for export.     

Divalent cations directly affect the conductance of excised patches of rod photoreceptor membrane

Phototransduction in rod cells is likely to involve an intracellular messenger system that links the absorption of light by rhodopsin to a change in membrane conductance. The direct effect of guanosine 3',5'-monophosphate (cGMP) on excised patches of rod outer segment membrane strongly supports a role for cGMP as an intracellular messenger in phototransduction. It is reported here that magnesium and calcium directly affect the conductance of excised patches of rod membrane in the absence of cGMP and that magnesium, applied to intact rod cells, blocks a component of the cellular light response. The divalent cation-suppressed conductance in excised patches showed outward rectification and cation-selective permeability resembling those of the light-suppressed conductance measured from the intact rod cell. The divalent cation-suppressed conductance was partly blocked by a concentration of the pharmacological agent L-cis-diltiazem that blocked all of the cGMP-activated conductance. Divalent cations may act, together with cGMP, as an intracellular messenger system that mediates the light response of the rod photoreceptor cell.     

Study on the Ultrastructure and its Change of Eggshell of Chukar during Incubation

Chukar is a kind of very important wild economical bird species.The research to the eggshell is very useful not only in biology but also in economics.The structure of eggshell of Chukar is the same as the fowl.The structural changes of the inner and outer surfaces,the transverse section of the eggshell,and the outer surface of its membrane during incubation had been observed by scanning electron microscopy (SEM).There was a great change happened for the surface structrue of eggshell and its membrane during incubation.The tortoise-shaped crackles became smaller and longer,and the fibres of the membrane became thinner and shorter.At the same time,the transverse section of eggshell also became thicker.     

The function of a leader peptide in translocating charged amino acyl residues across a membrane

Insertion of bacteriophage coat proteins into the membrane of infected bacterial cells can be studied as a model system of protein translocation across membranes. The coat protein of the filamentous bacteriophage Pf3--which infects Pseudomonas aeruginosa--is 44 amino acids in length and has the same basic structure as the coat protein of bacteriophage M13, which infects Escherichia coli. However, unlike the Pf3 coat protein, the M13 coat protein is synthesized as a precursor (procoat) with a typical leader (signal) sequence, which is cleaved after membrane insertion. Nevertheless, when the gene encoding the Pf3 coat protein is expressed in E. coli, the protein is translocated across the membrane. Hybrid M13 and Pf3 coat proteins were constructed in an attempt to understand how the Pf3 coat protein is translocated without a leader sequence. These studies demonstrated that the extracellular regions of the proteins determined their cellular location. When three charged residues in this region were neutralized, the leader-free M13 coat protein was also inserted into the membrane. Differences in the water shell surrounding these residues may account for efficient membrane insertion of the protein without a leader sequence.     

抗沙眼衣原体主要外膜蛋白(MOMP)单克隆抗体的制备及初步应用

制备抗沙眼衣原体(CT)单克隆抗体,建立沙眼衣原体的胶体金免疫层析快速检测方法。采用纯化沙眼衣原体主要外膜蛋白(MOMP)免疫Balb/c小鼠,选取高效价的免疫小鼠脾细胞与骨髓瘤细胞融合,ELISA检测筛选分泌抗MOMP单克隆抗体(McAb)的细胞株并进行相关鉴定。共获得分泌IgG1,IgG2b和IgM的3株单克隆抗体杂交瘤细胞株。纯化的单克隆抗体与鹦鹉热衣原体、肺炎衣原体、肺炎支原体均无交叉反应。通过位点分析选择配对良好的两株McAb,制备检测CT的双抗体夹心法免疫层析试纸条。制备的胶体金试纸条对MOMP的最低检出值可达50 ng·mL-1。     

丝状真菌遗传转化系统研究进展

近年来,丝状真菌的分子生物学研究发展非常迅速,尤其是丝状真菌遗传转化研究取得了长足的进展。对丝状真菌遗传转化系统的最新研究进展进行综述,主要包括丝状真菌的转化方法、选择标记、转化系统的应用等。     

动物双歧杆菌AD011暴露外表面蛋白的种类和功能分析

[目的]分析动物双歧杆菌AD011暴露外表面蛋白的种类和功能,有助于了解该菌株的各项生理功能以及为探讨该菌与宿主相互作用的分子机制奠定基础。[方法]以动物双歧杆菌AD011基因组序列为研究对象,采用Inmembrane方法和COG功能数据库对预测的外表面蛋白进行功能注释和聚类分析。[结果]AD011菌体暴露外表面蛋白数目为193个,包括11个细胞壁蛋白、20个脂蛋白、暴露外表面的膜蛋白140个、22个细胞外蛋白。AD011外表面蛋白的功能分析结果显示,193个外表面蛋白中,含有较大比例的蛋白没有功能注释(78个),115个蛋白具有COG功能注释。在有功能注释的蛋白中,所占比例较大的是氨基酸转运与代谢(16个),无机离子转运与代谢(14个),细胞壁、细胞膜生物合成(13个),碳水化合物转运与代谢(10个),防御机制(10个)等有关蛋白。[结论]AD011菌体外表面蛋白与营养物质的吸收和转运,细胞壁、细胞膜生物合成,防御机制有关。