Molecular basis of transmembrane signalling by sensory rhodopsin II-transducer complex

Microbial rhodopsins, which constitute a family of seven-helix membrane proteins with retinal as a prosthetic group, are distributed throughout the Bacteria, Archaea and Eukaryota. This family of photoactive proteins uses a common structural design for two distinct functions: light-driven ion transport and phototaxis. The sensors activate a signal transduction chain similar to that of the two-component system of eubacterial chemotaxis. The link between the photoreceptor and the following cytoplasmic signal cascade is formed by a transducer molecule that binds tightly and specifically to its cognate receptor by means of two transmembrane helices (TM1 and TM2). It is thought that light excitation of sensory rhodopsin II from Natronobacterium pharaonis (SRII) in complex with its transducer (HtrII) induces an outward movement of its helix F (ref. 6), which in turn triggers a rotation of TM2 (ref. 7). It is unclear how this TM2 transition is converted into a cellular signal. Here we present the X-ray structure of the complex between N. pharaonis SRII and the receptor-binding domain of HtrII at 1.94 A resolution, which provides an atomic picture of the first signal transduction step. Our results provide evidence for a common mechanism for this process in phototaxis and chemotaxis.     

平均配位数法求取蛋白质等电点

将蛋白质视为可连续电离出氢离子的多元弱酸,利用配合物体系配离子分布系数取最大值的条件-n=c,即蛋白质体系的平均配位数-n等于蛋白质中性离子中含有氢离子数目c时对应的pH值即为蛋白质等电点pI值,通过绘制蛋白质平均配位数-n对pH图或使用常用办公软件均可方便地找出pI值.考虑到蛋白质一般含有较多种类的由可电离游离基团电离形成的蛋白质离子,文章提出了忽略浓度较小蛋白质离子并简化-n计算的方法.     

单训练样本视频监控俯视人脸识别研究

近年来视频监控已普遍应用于各行各业,因此基于监控视频人脸识别也成为了智能监控系统中重要的研究领域.然而,由于监控视频人脸通常是非正面人脸,传统性能优良算法应用于视频人脸识别时,其性能也明显降低.同时,单张训练人脸问题在监控视频人脸检测和识别是一个普遍问题.因此为了能有效地提高单训练多姿态人脸识别的正确识别率,文章提出了一种基于三维建模技术的人脸识别算法.该算法先由一张二维高清正面人脸生成一个三维人脸模型,然后再进一步在该三维人脸空间里产生多种姿态的人脸模型,并由此获得多张相应姿态下的二维虚拟人脸,最后利用原始正面样本和所得到的虚拟人脸来构筑训练人脸库.该算法用SCface视频监控人脸库中加以验证,与传统的PCA和LDA算法相比,该算法对监控视频人脸的识别率提高了13%.由此表明,文章介绍的算法是一种有效的人脸识别算法,能有效地提高对俯视人脸的识别率.     

Leukocytes express a novel gene encoding a putative transmembrane protein-kinase devoid of an extracellular domain

Tyrosine-specific phosphorylation of proteins is a key to the control of diverse pathways leading to cell growth and differentiation. The protein-tyrosine kinases described to date are either transmembrane proteins having an extracellular ligand binding domain or cytoplasmic proteins related to the v-src oncogene. Most of these proteins are expressed in a wide variety of cells and tissues; few are tissue-specific. Previous studies have suggested that lymphokines could mediate haematopoietic cell survival through their action on glucose transport, regulated in some cells through the protein-tyrosine kinase activity of the insulin receptor. We have investigated the possibility that insulin receptor-like genes are expressed specifically in haematopoietic cells. Using the insulin receptor-related avian sarcoma oncogene v-ros as a probe, we have isolated and characterized the complementary DNA of a novel gene, ltk (leukocyte tyrosine kinase). The ltk gene is expressed mainly in leukocytes, is related to several tyrosine kinase receptor genes of the insulin receptor family and has unique structural properties: it apparently encodes a transmembrane protein devoid of an extracellular domain. Two candidate ltk proteins have been identified with antibodies in the mouse thymus, and have properties indicating that they are integral membrane proteins. These features suggest that ltk could be a signal transduction subunit for one or several of the haematopoietic receptors.     

Enhanced partner preference in a promiscuous species by manipulating the expression of a single gene

The molecular mechanisms underlying the evolution of complex behaviour are poorly understood. The mammalian genus Microtus provides an excellent model for investigating the evolution of social behaviour. Prairie voles (Microtus ochrogaster) exhibit a monogamous social structure in nature, whereas closely related meadow voles (Microtus pennsylvanicus) are solitary and polygamous. In male prairie voles, both vasopressin and dopamine act in the ventral forebrain to regulate selective affiliation between adult mates, known as pair bond formation, as assessed by partner preference in the laboratory. The vasopressin V1a receptor (V1aR) is expressed at higher levels in the ventral forebrain of monogamous than in promiscuous vole species, whereas dopamine receptor distribution is relatively conserved between species. Here we substantially increase partner preference formation in the socially promiscuous meadow vole by using viral vector V1aR gene transfer into the ventral forebrain. We show that a change in the expression of a single gene in the larger context of pre-existing genetic and neural circuits can profoundly alter social behaviour, providing a potential molecular mechanism for the rapid evolution of complex social behaviour.     

Structural basis for Duffy recognition by the malaria parasite Duffy-binding-like domain

Molecular processes that govern pathogenic features of erythrocyte invasion and cytoadherence in malaria are reliant on Plasmodium-specific Duffy-binding-like domains (DBLs). These cysteine-rich modules recognize diverse host cell-surface receptors during pathogenesis. DBLs of parasite erythrocyte-binding proteins mediate invasion, and those from the antigenically variant P. falciparum erythrocyte membrane protein 1 (PfEMP1) have been implicated in cytoadherence. The simian and human malarial parasites, P. knowlesi and P. vivax, invade human erythrocytes exclusively through the host DARC receptor (Duffy antigen receptor for chemokines). Here we present the crystal structure of the P. knowlesi DBL domain (Pkalpha-DBL), which binds to DARC during invasion of human erythrocytes. Pkalpha-DBL retains the overall fold observed in DBLs from P. falciparum erythrocyte-binding antigen (EBA)-175 (ref. 4). Mapping the residues that have previously been implicated in binding highlights a fairly flat but exposed site for DARC recognition in subdomain 2 of Pkalpha-DBL; this is in sharp contrast to receptor recognition by EBA-175 (ref. 4). In Pkalpha-DBL, the residues that contact DARC and the clusters of residues under immune pressure map to opposite surfaces of the DBL, and suggest a possible mechanism for immune evasion by P. vivax. Our comparative structural analysis of Pkalpha-DBL and P. falciparum EBA-175 provides a framework for the understanding of malaria parasite DBLs, and may affect the development of new prophylactic and therapeutic strategies.     

Plasma and cytoplasmic gelsolins are encoded by a single gene and contain a duplicated actin-binding domain

Gelsolin is representative of a class of actin-modulating proteins found in lower eukaryotes to mammals, which sever actin filaments. Gelsolin found in the cytoplasm of cells is functionally similar to a mammalian plasma protein of similar size, originally called ADF or brevin. Human plasma and rabbit macrophage gelsolins differ by the presence of a 25-amino-acid residue extension on plasma gelsolin which appears to account for the difference in relative molecular mass (Mr) between the proteins as assessed by SDS-polyacrylamide gel electrophoresis (PAGE), 93,000 (93K) and 90K, respectively. Here we report the isolation of full-length human plasma gelsolin complementary DNA clones from a HepG2 library. The inferred amino-acid sequence reveals the presence of a signal peptide, a long tandem repeat that matches the actin-binding domains of gelsolin, a tetrapeptide present in actin and extended regions of identical sequence with rabbit macrophage gelsolin. Southern blot analysis indicates that a single gene in the haploid genome encodes both protein forms.     

增长区域上两种群的竞争和扩散

考虑了增长区域上一类具扩散的Lotka-Volterra竞争模型,定义了生态再生指标并将其作为阈值。探讨了模型的稳定态以及初边值问题解的渐近行为。理论结果与数值模拟均表明,生存区域的增长对竞争种群的双方都有利。     

Current-induced resonance and mass determination of a single magnetic domain wall

A magnetic domain wall (DW) is a spatially localized change of magnetization configuration in a magnet. This topological object has been predicted to behave at low energy as a composite particle with finite mass. This particle will couple directly with electric currents as well as magnetic fields, and its manipulation using electric currents is of particular interest with regard to the development of high-density magnetic memories. The DW mass sets the ultimate operation speed of these devices, but has yet to be determined experimentally. Here we report the direct observation of the dynamics of a single DW in a ferromagnetic nanowire, which demonstrates that such a topological particle has a very small but finite mass of 6.6 x 10(-23) kg. This measurement was realized by preparing a tunable DW potential in the nanowire, and detecting the resonance motion of the DW induced by an oscillating current. The resonance also allows low-current operation, which is crucial in device applications; a DW displacement of 10 microm was induced by a current density of 10(10) A m(-2).     

Structural basis for overhang-specific small interfering RNA recognition by the PAZ domain

Short RNAs mediate gene silencing, a process associated with virus resistance, developmental control and heterochromatin formation in eukaryotes. RNA silencing is initiated through Dicer-mediated processing of double-stranded RNA into small interfering RNA (siRNA). The siRNA guide strand associates with the Argonaute protein in silencing effector complexes, recognizes complementary sequences and targets them for silencing. The PAZ domain is an RNA-binding module found in Argonaute and some Dicer proteins and its structure has been determined in the free state. Here, we report the 2.6 A crystal structure of the PAZ domain from human Argonaute eIF2c1 bound to both ends of a 9-mer siRNA-like duplex. In a sequence-independent manner, PAZ anchors the 2-nucleotide 3' overhang of the siRNA-like duplex within a highly conserved binding pocket, and secures the duplex by binding the 7-nucleotide phosphodiester backbone of the overhang-containing strand and capping the 5'-terminal residue of the complementary strand. On the basis of the structure and on binding assays, we propose that PAZ might serve as an siRNA-end-binding module for siRNA transfer in the RNA silencing pathway, and as an anchoring site for the 3' end of guide RNA within silencing effector complexes.     

采用顶部籽晶熔融辅助熔渗法制备高性能单畴GdBCO超导块材

在顶部籽晶熔渗生长工艺(TSIG)的基础上,本文采用熔融织构生长(MTG)辅助法制备高性能大尺寸单畴GdBCO超导块材,并且对样品的生长形貌、晶体界面生长结构、磁悬浮力以及捕获磁通进行了研究分析.研究结果表明,采用顶部籽晶熔融辅助熔渗生长工艺可以有效地提高NdBCO籽晶的利用率,抑制样品在生长过程中产生随机成核现象,从而大大地提高制备单畴样品的有效率.此外,用新的生长工艺还有助于提高GdBCO样品的性能.     

Molecular recognition study on a supramolecular system (VIII)

The fluorescence spectral titrations have been performed in phosphate buffer solution (pH 7.2) at room temperature to give the stability constants for inclusion complexation of a series of chemically modified β-cyclodextrins bearing aromatic groups with adamantanol. Thc molecular recognition mechanism is discussed from the viewpoint of the structural factors, electron effect of the substitutent groups and thc size and chain length of the fluorophore. The results indicate that the complex stability of the cyclodextrin derivatives bearing a positively charged pyridinio moiety can be enhanced by the electrostatic interaction between the host cyclodextrins and the adamantanol guest and the microenvironment change plays a crucial role in the inclusion complexation.     

Molecular recognition study on a supramolecular system (XV)

The spectrophotometric titrations have been performed at 298.15 K in buffered aqueous solution (pH 7.20) to give the stability constants (logK₅) for the supramolecular system formed by mono-[6-(phenylseleno)-6-deoxy]-β-cyclodextrin (2), mono-[6-(p-tolylseleno)-6-deoxy]-β-cyclodextrin (3), mono[6-(o-tolylseleno)-6-deoxy]-β-cyclodextrin (4), mono-[6-(m-tolylseleno)-6-deoxy]-β-cyclodextrin (5) with a series of aliphatic alcohols. The results were compared with the data reported for the parent β-cyclodextrin. The molecular binding ability and selectivity for aliphatic alcohols of modified β-cyclodextrins (2–5) were discussed from the viewpoints of the induced-fit and geometrical complement. The results obtained indicate that the phenylseleno moiety of β-cyclodextrin derivatives (2–5) as a probe for spectrometry can enhance not only the molecular binding ability but also the molecular recognition ability.     

一个离散单种群扩散模型的全局渐近稳定性

建立了一个物种在ｎ个缀块上相互扩散的数学模型并利用单调算子和凹算子的理论讨论了其全局渐近稳定性,得到了正平衡点全局渐近稳定和物种绝灭的条件．     

Molecular dynamics simulation of a single polymer in hydrophilic nano-slits

The behavior of a single polyethylene polymer in aqueous solution confined between two hydrophilic walls is studied with molecular dynamics （MD） simulations. The thickness of the nano-slit ranges from 1.26 to 3.15 nm, which is comparative to the polymer dimension. A monotonic transition from 3D- to 2D-like configurations is observed as the distance between the two walls narrows. Monomers are compressed into several layers and the preferred bond orientations alternate between parallel and normal to the walls accordingly. The diffusivity in the direction parallel to the wall is always larger than the one perpendicular to it. Calculation of the entropy and enthalpy changes during the folding of the polymer chain alone cannot explain the spontaneous process. The corresponding increase in water entropy due to volume expansion may be large enough to result in the overall free energy decrease.     

Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases

Xeroderma pigmentosum is a monogenic disease characterized by hypersensitivity to ultraviolet light. The cells of xeroderma pigmentosum patients are defective in nucleotide excision repair, limiting their capacity to eliminate ultraviolet-induced DNA damage, and resulting in a strong predisposition to develop skin cancers. The use of rare cutting DNA endonucleases-such as homing endonucleases, also known as meganucleases-constitutes one possible strategy for repairing DNA lesions. Homing endonucleases have emerged as highly specific molecular scalpels that recognize and cleave DNA sites, promoting efficient homologous gene targeting through double-strand-break-induced homologous recombination. Here we describe two engineered heterodimeric derivatives of the homing endonuclease I-CreI, produced by a semi-rational approach. These two molecules-Amel3-Amel4 and Ini3-Ini4-cleave DNA from the human XPC gene (xeroderma pigmentosum group C), in vitro and in vivo. Crystal structures of the I-CreI variants complexed with intact and cleaved XPC target DNA suggest that the mechanism of DNA recognition and cleavage by the engineered homing endonucleases is similar to that of the wild-type I-CreI. Furthermore, these derivatives induced high levels of specific gene targeting in mammalian cells while displaying no obvious genotoxicity. Thus, homing endonucleases can be designed to recognize and cleave the DNA sequences of specific genes, opening up new possibilities for genome engineering and gene therapy in xeroderma pigmentosum patients whose illness can be treated ex vivo.     

Chi-sequence recognition and DNA translocation by single RecBCD helicase/nuclease molecules

Major pathways of recombinational DNA repair in Escherichia coli require the RecBCD protein--a heterotrimeric, ATP-driven, DNA translocating motor enzyme. RecBCD combines a highly processive and exceptionally fast helicase (DNA-unwinding) activity with a strand-specific nuclease (DNA-cleaving) activity (refs 1, 2 and references therein). Recognition of the DNA sequence 'chi' (5'-GCTGGTGG-3') switches the polarity of DNA cleavage and stimulates recombination at nearby sequences in vivo. Here we attach microscopic polystyrene beads to biotin-tagged RecD protein subunits and use tethered-particle light microscopy to observe translocation of single RecBCD molecules (with a precision of up to approximately 30 nm at 2 Hz) and to examine the mechanism by which chi modifies enzyme activity. Observed translocation is unidirectional, with each molecule moving at a constant velocity corresponding to the population-average DNA unwinding rate. These observations place strong constraints on possible movement mechanisms. Bead release at chi is negligible, showing that the activity modification at chi does not require ejection of the RecD subunit from the enzyme as previously proposed; modification may occur through an unusual, pure conformational switch mechanism.     

Structural basis for recognition of hemi-methylated DNA by the SRA domain of human UHRF1

Epigenetic inheritance in mammals is characterized by high-fidelity replication of CpG methylation patterns during development. UHRF1 (also known as ICBP90 in humans and Np95 in mouse) is an E3 ligase important for the maintenance of global and local DNA methylation in vivo. The preferential affinity of UHRF1 for hemi-methylated DNA over symmetrically methylated DNA by means of its SET and RING-associated (SRA) domain and its association with the maintenance DNA methyltransferase 1 (DNMT1) suggests a role in replication of the epigenetic code. Here we report the 1.7 A crystal structure of the apo SRA domain of human UHRF1 and a 2.2 A structure of its complex with hemi-methylated DNA, revealing a previously unknown reading mechanism for methylated CpG sites (mCpG). The SRA-DNA complex has several notable structural features including a binding pocket that accommodates the 5-methylcytosine that is flipped out of the duplex DNA. Two specialized loops reach through the resulting gap in the DNA from both the major and the minor grooves to read the other three bases of the CpG duplex. The major groove loop confers both specificity for the CpG dinucleotide and discrimination against methylation of deoxycytidine of the complementary strand. The structure, along with mutagenesis data, suggests how UHRF1 acts as a key factor for DNMT1 maintenance methylation through recognition of a fundamental unit of epigenetic inheritance, mCpG.