Crystal structure of metarhodopsin II

G-protein-coupled receptors (GPCRs) are seven transmembrane helix (TM) proteins that transduce signals into living cells by binding extracellular ligands and coupling to intracellular heterotrimeric G proteins (Gαβγ). The photoreceptor rhodopsin couples to transducin and bears its ligand 11-cis-retinal covalently bound via a protonated Schiff base to the opsin apoprotein. Absorption of a photon causes retinal cis/trans isomerization and generates the agonist all-trans-retinal in situ. After early photoproducts, the active G-protein-binding intermediate metarhodopsin II (Meta?II) is formed, in which the retinal Schiff base is still intact but deprotonated. Dissociation of the proton from the Schiff base breaks a major constraint in the protein and enables further activating steps, including an outward tilt of TM6 and formation of a large cytoplasmic crevice for uptake of the interacting C terminus of the Gα subunit. Owing to Schiff base hydrolysis, Meta?II is short-lived and notoriously difficult to crystallize. We therefore soaked opsin crystals with all-trans-retinal to form Meta?II, presuming that the crystal's high concentration of opsin in an active conformation (Ops*) may facilitate all-trans-retinal uptake and Schiff base formation. Here we present the 3.0?? and 2.85?? crystal structures, respectively, of Meta?II alone or in complex with an 11-amino-acid C-terminal fragment derived from Gα (GαCT2). GαCT2 binds in a large crevice at the cytoplasmic side, akin to the binding of a similar Gα-derived peptide to Ops* (ref. 7). In the Meta?II structures, the electron density from the retinal ligand seamlessly continues into the Lys?296 side chain, reflecting proper formation of the Schiff base linkage. The retinal is in a relaxed conformation and almost undistorted compared with pure crystalline all-trans-retinal. By comparison with early photoproducts we propose how retinal translocation and rotation induce the gross conformational changes characteristic for Meta?II. The structures can now serve as models for the large GPCR family.     

MiR1511 co-regulates with miR1511* to cleave the GmRPL4a gene in soybean

MicroRNA1511 (miR1511) is a small RNA with unknown function identified in several plants by deep sequencing. In this study, we showed that this small RNA is an authentic miRNA by analyzing the structure of the precursor stem-loop containing the newly identified miR1511* sequence. We confirmed this result by Northern blotting analysis. We used 5??RACE to identify one of the target genes (GmRPL4a) cleaved by both miR1511 and miR1511*. The site cleaved by miR1511* was located in the first exon of GmRPL4a, and the site cleaved by miR1511 was located in the second exon. The expression level of miR1511/1511* was higher in leaves than in roots and stems. In contrast, the lowest level of GmRPL4a expression was in the leaves and the highest in the root. These results indicate that an miRNA can co-regulate with an miRNA* to cleave the same target gene in plants, and that the level of GmRPL4a mRNA is regulated by miR1511/1511*.     

Crystal structure of the β2 adrenergic receptor-Gs protein complex

G protein-coupled receptors (GPCRs) are responsible for the majority of cellular responses to hormones and neurotransmitters as well as the senses of sight, olfaction and taste. The paradigm of GPCR signalling is the activation of a heterotrimeric GTP binding protein (G protein) by an agonist-occupied receptor. The β(2) adrenergic receptor (β(2)AR) activation of Gs, the stimulatory G protein for adenylyl cyclase, has long been a model system for GPCR signalling. Here we present the crystal structure of the active state ternary complex composed of agonist-occupied monomeric β(2)AR and nucleotide-free Gs heterotrimer. The principal interactions between the β(2)AR and Gs involve the amino- and carboxy-terminal α-helices of Gs, with conformational changes propagating to the nucleotide-binding pocket. The largest conformational changes in the β(2)AR include a 14 ? outward movement at the cytoplasmic end of transmembrane segment 6 (TM6) and an α-helical extension of the cytoplasmic end of TM5. The most surprising observation is a major displacement of the α-helical domain of Gαs relative to the Ras-like GTPase domain. This crystal structure represents the first high-resolution view of transmembrane signalling by a GPCR.     

Crystal structure of the ligand-free G-protein-coupled receptor opsin

In the G-protein-coupled receptor (GPCR) rhodopsin, the inactivating ligand 11-cis-retinal is bound in the seven-transmembrane helix (TM) bundle and is cis/trans isomerized by light to form active metarhodopsin II. With metarhodopsin II decay, all-trans-retinal is released, and opsin is reloaded with new 11-cis-retinal. Here we present the crystal structure of ligand-free native opsin from bovine retinal rod cells at 2.9 ?ngstr?m (A) resolution. Compared to rhodopsin, opsin shows prominent structural changes in the conserved E(D)RY and NPxxY(x)(5,6)F regions and in TM5-TM7. At the cytoplasmic side, TM6 is tilted outwards by 6-7 A, whereas the helix structure of TM5 is more elongated and close to TM6. These structural changes, some of which were attributed to an active GPCR state, reorganize the empty retinal-binding pocket to disclose two openings that may serve the entry and exit of retinal. The opsin structure sheds new light on ligand binding to GPCRs and on GPCR activation.     

C_PCP与_w_w_CPCP的关系

本文证明了下面２个结果：（１）当Ｘ＊具有ＰＣＰ时，Ｃ＊ＰＣＰ与（ｗ＊－ｗ）ＣＰＣＰ等价；（２）Ｘ＊具有Ｃ＊ＰＣＰ当且仅当Ｘ＊具有（ｗ＊－ｗ）ＣＰＣＰ且对Ｘ＊的每个非空ｗ＊－紧凸子集Ｋ，（ｗ＊－ｗ）ＰＣ（Ｋ）含有Ｋ的一个ｗ＊－稠密Ｇ子集．     

极小3连通图的非基本边数

利用扇,断片及简约图的概念,得到不为轮的极小3连通图的非基本边数与其简约图的非基本边数相等,从而将求极小3连通图的非基本边数问题转化为求其简约图的非基本边数问题后,给出简约极小3连通图非基本边数的一个下界,刻画了达到下界的图类.     

3,6-二甲氧基环丙基萘的结构和性质研究

在RHF/6 31G 和B3LYP/6 31G 水平下优化了3,6 二甲氧基环丙基萘(MOCPN)的平均几何构型,用B3LYP/6 31G 方法计算了该化合物的红外光谱,并用GIAO分别在B3LYP/6 31G 、B3LYP/6 311G 和B3LYP/6 311 G 水平对该化合物的核磁共振谱进行了研究;计算结果与实验结果吻合很好.     

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.     

Towards a Full-Stack Dev Ops Environment (Platform-as-a-Service) for Cloud-Hosted Applications

If every programmer of cloud-hosted apps possessed exceptional technical capability and endless patience, the Dev Ops environment(also known as Platform-as-a-Service, or Paa S) would perhaps become irrelevant. However, the reality is almost always the opposite case. Hence, IT engineers dream of a reliable and usable Dev Ops environment that can substantially facilitate their developments and simplify their operations.Current Dev Ops environments include Google App Engine, Docker, Kubernetes, Mesos, and so forth. In other words, Paa S bridges the gap between vivid IT engineers and stiff cloud systems. In this paper, we comprehensively examine state-of-the-art Paa S solutions across various tiers of the cloud-computing Dev Ops stack. On this basis,we identify areas of consensus and diversity in their philosophies and methodologies. In addition, we explore cutting-edge solutions towards realizing a more fine-grained, full-stack Dev Ops environment. From this paper,readers are expected to quickly grasp the essence, current status, and future prospects of Paa S.     

Role of a subdomain in the folding of bovine pancreatic trypsin inhibitor.

The disulphide-bonded intermediates that accumulate in the oxidative folding of bovine pancreatic trypsin inhibitor (BPTI) were characterized some time ago. Structural characterization of these intermediates would provide an explanation of the kinetically preferred pathways of folding for BPTI. When folding occurs under strongly oxidizing conditions, more than half the molecules become trapped in an intermediate, designated N*, which is similar to the native protein but lacks the 30-51 disulphide bond. We have tested the hypothesis that the precursor to N* is the one-disulphide intermediate [5-55], which contains the most stable disulphide in BPTI, and present evidence here that this is the case. A peptide model of [5-55], corresponding to a subdomain of BPTI, seems to fold into a native-like conformation, explaining why [5-55] does not lead to native protein and why it folds rapidly to N*. A native-like subdomain structure in a peptide model of [30-51], the other crucial one-disulphide intermediate, may explain the route by which [30-51] folds to native protein. Thus, much of the folding pathway of BPTI can be explained by the formation of a native-like subdomain in these two early intermediates. This suggests that a large part of the protein folding problem can be reduced to identifying and understanding subdomains of native proteins.     

Cis-trans isomerization at a proline opens the pore of a neurotransmitter-gated ion channel

5-hydroxytryptamine type 3 (5-HT3) receptors are members of the Cys-loop receptor superfamily. Neurotransmitter binding in these proteins triggers the opening (gating) of an ion channel by means of an as-yet-uncharacterized conformational change. Here we show that a specific proline (Pro 8*), located at the apex of the loop between the second and third transmembrane helices (M2-M3), can link binding to gating through a cis-trans isomerization of the protein backbone. Using unnatural amino acid mutagenesis, a series of proline analogues with varying preference for the cis conformer was incorporated at the 8* position. Proline analogues that strongly favour the trans conformer produced non-functional channels. Among the functional mutants there was a strong correlation between the intrinsic cis-trans energy gap of the proline analogue and the activation of the channel, suggesting that cis-trans isomerization of this single proline provides the switch that interconverts the open and closed states of the channel. Consistent with this proposal, nuclear magnetic resonance studies on an M2-M3 loop peptide reveal two distinct, structured forms. Our results thus confirm the structure of the M2-M3 loop and the critical role of Pro 8* in the 5-HT3 receptor. In addition, they suggest that a molecular rearrangement at Pro 8* is the structural mechanism that opens the receptor pore.     

对银河系中心超大质量黑洞候选体Sgr A*的VLBI观测研究

位于银河系中央的极其致密的非热射电源Sagittari8us A^*（Sgr A^*），被公认为最佳的超大质量黑洞候选体，其质量约为400万个太阳质量。甚长基线干涉测量（BLBI）技术以其独有的高空间分辩率为我们提供了唯一可以探讨Sgr A^*位于几个史瓦西（Schwarzschild）半径区域内天体物理性质的手段。介绍了对 Sgr A^*的VLBI研究现状和最新进展，着重论述了毫米波和亚毫米波VLBI观测对研究Sgr A^*的辐射区域的真实形状和大小，以及探索其结构随时间变化的重要性。Sgr A^*无疑是检验广义相对论将就的亚亮米波VLBI实验的最佳目标。     

标准C*-代数间保持一类正元比较关系的映射

假设H是一个复Hilbert空间,AH是H上的一个标准C*-代数,AH+是AH中所有正元构成的集合.1977年,Cuntz引入了C*-代数中正元的比较关系.设A是一个C*-代数,任给A,B∈A+,若存在X∈A+,使得A=XBX*,则记A≤B.文章刻画了定义在AH+上的所有双边保持此关系的弱连续半线性满射.     

连续参数集值下鞅的Doob-Meyer分解定理

给出了两个定理和两个推论,定理1为：若X^*可分为,y包含f为σ域,则F：Ω→Pfc(X)为y可测的充要条件为倒AX^*∈X^*,ω→（X^*,F(ω）为y可测的,定理2给出了连续参数值下鞅存在唯一的Doob-Meyer分解的充要条件。     

单调算子与伪单调算子和的值域

设X是实Banach空间,X*是其对偶空间.T:X( )D(T)→X*是单调算子,C:X( )D(T)→X*是伪单调算子,本文主要利用S+型算子的度理论讨论了当X是自反Banach空间且X和X*均为局部一致凸空间时单调与伪单调算子和的值域问题,在证明过程中主要应用了伪单调算子与S+型算子的和仍是S+型算子这一理论.     

Structure of the nociceptin/orphanin FQ receptor in complex with a peptide mimetic

Members of the opioid receptor family of G-protein-coupled receptors (GPCRs) are found throughout the peripheral and central nervous system, where they have key roles in nociception and analgesia. Unlike the 'classical' opioid receptors, δ, κ and μ (δ-OR, κ-OR and μ-OR), which were delineated by pharmacological criteria in the 1970s and 1980s, the nociceptin/orphanin FQ (N/OFQ) peptide receptor (NOP, also known as ORL-1) was discovered relatively recently by molecular cloning and characterization of an orphan GPCR. Although it shares high sequence similarity with classical opioid GPCR subtypes (～60%), NOP has a markedly distinct pharmacology, featuring activation by the endogenous peptide N/OFQ, and unique selectivity for exogenous ligands. Here we report the crystal structure of human NOP, solved in complex with the peptide mimetic antagonist compound-24 (C-24) (ref. 4), revealing atomic details of ligand-receptor recognition and selectivity. Compound-24 mimics the first four amino-terminal residues of the NOP-selective peptide antagonist UFP-101, a close derivative of N/OFQ, and provides important clues to the binding of these peptides. The X-ray structure also shows substantial conformational differences in the pocket regions between NOP and the classical opioid receptors κ (ref. 5) and μ (ref. 6), and these are probably due to a small number of residues that vary between these receptors. The NOP-compound-24 structure explains the divergent selectivity profile of NOP and provides a new structural template for the design of NOP ligands.     

视网膜振荡电位异常改变的临床分析

目的 对多种眼科疾病进行视网膜振荡电位(Ops)的检测及分析，探讨了视网膜振荡电位在眼科多种疾病的诊断及评估方面的临床意义。方法对261例412只眼进行Ops检测，对Ops的检测结果进行分析。结果发现糖尿病及糖尿病性视网膜病变、视网膜中央及分支静脉阻塞、高度近视及高度近视性视网膜病变、视网膜色素变性等均有不同程度的Ops总幅值改变。同时发现，老年性黄斑变性、老年性白内障、弱视、中心性浆液性视网膜脉络膜病变及轻、中度近视者，则Ops总幅值无明显改变。结论视网膜Ops的检测是临床眼科诊断及评估的有力手段。     

一族由前缀码生成的极大自由幺子半群

设X*是由字母表X生成的自由幺半群且A是X*的非空子集,如果A∩AX+=Φ,则称A是前缀码。设{B1,B2}是X的任意2—划分,令A=B2∪B1(Xi\Bi1)∪E,i=1,2,其中E=Bi1+1(B01B1∪B2B1∪B22B1∪…∪B2M-1B1∪B2MX),M≥0。文章证明了A是前缀码且幺半群A*是自由幺半群X*的极大自由幺子半群。     

丹参醌类抗氧化剂的结构特征与活性关联(Ⅰ)

对双氢丹参醌Ⅰ和丹参醌Ⅱ在油脂中的抗氧化作用及构效关联进行了量子化学研究.结果表明，抗氧化剂与油脂中碳中心自由基（R