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 oxygen-evolving photosystem II at a resolution of 1.9 Å

Photosystem II is the site of photosynthetic water oxidation and contains 20 subunits with a total molecular mass of 350 kDa. The structure of photosystem II has been reported at resolutions from 3.8 to 2.9 ?. These resolutions have provided much information on the arrangement of protein subunits and cofactors but are insufficient to reveal the detailed structure of the catalytic centre of water splitting. Here we report the crystal structure of photosystem II at a resolution of 1.9 ?. From our electron density map, we located all of the metal atoms of the Mn(4)CaO(5) cluster, together with all of their ligands. We found that five oxygen atoms served as oxo bridges linking the five metal atoms, and that four water molecules were bound to the Mn(4)CaO(5) cluster; some of them may therefore serve as substrates for dioxygen formation. We identified more than 1,300 water molecules in each photosystem II monomer. Some of them formed extensive hydrogen-bonding networks that may serve as channels for protons, water or oxygen molecules. The determination of the high-resolution structure of photosystem II will allow us to analyse and understand its functions in great detail.     

Molecular modeling of the ion channel-like nanotube structure of amyloid β-peptide

The ion channel-like nanotube structure of the oligomers of amyloid β-peptide (Aβ) was first investi-gated by molecular modeling. The results reveal that the hydrogen bond net is one of the key factors to stabilize the structure. The hydrophobicity distribution mode of the side chains is in favor of the structure inserting into the bilayers and forming a hydrophilic pore. The lumen space is under the control of the negative potential,weaker but spreading continuously,to which the cation selectivity attributes; meanwhile,the alternate distribution of the stronger positive and negative potentials makes the electrostatic distribution of the structure framework balance,which is also one of the key factors stabilizing the structure. The results lay the theoretical foundation for illuminating the structure stability and the ion permeability,and give a clue to elucidating the molecular mechanism of Alzheimer's dis-ease (AD) and designing novel drugs to prevent or reverse AD at the root.     

Crystal structure of the µ-opioid receptor bound to a morphinan antagonist

Opium is one of the world's oldest drugs, and its derivatives morphine and codeine are among the most used clinical drugs to relieve severe pain. These prototypical opioids produce analgesia as well as many undesirable side effects (sedation, apnoea and dependence) by binding to and activating the G-protein-coupled μ-opioid receptor (μ-OR) in the central nervous system. Here we describe the 2.8?? crystal structure of the mouse μ-OR in complex with an irreversible morphinan antagonist. Compared to the buried binding pocket observed in most G-protein-coupled receptors published so far, the morphinan ligand binds deeply within a large solvent-exposed pocket. Of particular interest, the μ-OR crystallizes as a two-fold symmetrical dimer through a four-helix bundle motif formed by transmembrane segments 5 and 6. These high-resolution insights into opioid receptor structure will enable the application of structure-based approaches to develop better drugs for the management of pain and addiction.     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria (AOB) in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism (RFLP) analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the Nitrosomonas marina and the Nitrosomonas sp. Nm 143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

Crystal structure of a concentrative nucleoside transporter from Vibrio cholerae at 2.4 Å

Nucleosides are required for DNA and RNA synthesis, and the nucleoside adenosine has a function in a variety of signalling processes. Transport of nucleosides across cell membranes provides the major source of nucleosides in many cell types and is also responsible for the termination of adenosine signalling. As a result of their hydrophilic nature, nucleosides require a specialized class of integral membrane proteins, known as nucleoside transporters (NTs), for specific transport across cell membranes. In addition to nucleosides, NTs are important determinants for the transport of nucleoside-derived drugs across cell membranes. A wide range of nucleoside-derived drugs, including anticancer drugs (such as Ara-C and gemcitabine) and antiviral drugs (such as zidovudine and ribavirin), have been shown to depend, at least in part, on NTs for transport across cell membranes. Concentrative nucleoside transporters, members of the solute carrier transporter superfamily SLC28, use an ion gradient in the active transport of both nucleosides and nucleoside-derived drugs against their chemical gradients. The structural basis for selective ion-coupled nucleoside transport by concentrative nucleoside transporters is unknown. Here we present the crystal structure of a concentrative nucleoside transporter from Vibrio cholerae in complex with uridine at 2.4??. Our functional data show that, like its human orthologues, the transporter uses a sodium-ion gradient for nucleoside transport. The structure reveals the overall architecture of this class of transporter, unravels the molecular determinants for nucleoside and sodium binding, and provides a framework for understanding the mechanism of nucleoside and nucleoside drug transport across cell membranes.     

Crystal structure and surface photo-electric property of Mn（Ⅱ） coordination supramolecules

Two Mn（Ⅱ） coordination supramolecules, [Mn2（C8HTO2）4（phen）2（p-H20）] （1） and [Mn2（btec）（phen）2（H2O）6]·2H2O （2） （phen=1,10-phenanthroline, H4btec=1,2,4,5-benzenetetracarboxylic acid）, were synthesized by hydrothermal method. The crystal structures of the complexes were determined by X-ray single crystal diffraction. The result indicates that （1） and （2） are both binuclear Mn（Ⅱ） complexes. The existence of hydrogen bonds makes the binuclear complexes become further connected to coordination supramolecules, which possess 1D and 3D infinite structures respectively. The complexes were identified by IR, UV-Vis, surface photovoltage spectrum （SPS） and field-induced surface photovoltage spectrum （FISPS）. The results of SPS for the complexes indicate that they both exhibit positive surface photovoltage response bands in the range of 300-600 nm. The SPS phase spectrum and FISPS of complexes indicate that they show certain p-type semiconductor characteristic. However, the intensity, position and number of the SPV response bands are different obviously. The difference of the SPV response bands is mainly attributed to the different structures of the complexes and the different coordination environment of Mn（ll） in the two complexes. This paper discusses the action of hydrogen bonds in the construction of the supramolecule and the change on the surface photovoltage of complex in different coordination environment.     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria (AOB) in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism (RFLP) analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the N. marina and the N. sp. Nm143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

Community structure of β-Proteobacterial ammonia-oxidizing bacteria in prawn farm sediment

To examine the community structure of β-Proteobacterial ammonia-oxidizing bacteria （AOB） in prawn farm sediment, the 16S rRNA gene library was constructed with β-Proteobacterial AOB-specific primers. The library was screened by PCR-restriction fragment length polymorphism （RFLP） analysis and clones with unique RFLP patterns were sequenced. Two groups of β-Proteobacterial AOB, the Nitrosomonas and the Nitrosospira, were detected. The Nitrosomonas occupied an absolute dominant position, accounting for more than 90% of total clones in the clone library, while the Nitrosospira accounting for 5.48%. Nitrosomonas-affiliated clones were grouped into the Nitrosomonas marina and the Nitrosomonas sp. Nm143 clusters, and Nitrosospira-affiliated clones were grouped into the Nitrosospira cluster 1. No other clusters of β-Proteobacterial AOB were found. The results enriched our knowledge of AOB diversity in the prawn farm sediment and provided important foundational data for further functional studies of these microbes in mariculture environments.     

Crystal structure and photoelectricity property of Fe(Ⅱ/Ⅲ) coordination supermolecules

Three new mixed-ligand Fe(Ⅱ/Ⅲ) complexes [Fe2(μ2–btec)( μ2–H2btec)(phen)2(H2O)2]n (1), [Fe2(btec) (phen)2(H2O)4] (2), and {[Fe(o-pha)(phen)(H2O)]·H2O}n (3) (phen=1,10-phenanthroline, o-H2pha=o-phthalic acid, H4btec=1,2,4,5-benzenetetracarboxylic acid) have been hydrothermally synthesized and detected by single crystal X-ray diffraction, showing that complexes (1) and (2) are both bridged by the betc4? ligands to form 1D chain and dinuclear structure and complex (3) is bridged by the o-pha groups to form 1D ...     

The ring-substituted phthalo-cyanines and its metal com-plexes: Crystal structure of 1,4, 8,11,15,18,22,25-octa-butoxyphthalocyani-natocopper(Ⅱ)

Crystal structure of 1, 4, 8, 11, 15, 18, 22, 25-octa-butoxyphthalocyaninato- copper (Ⅱ) (1) was determined by X-ray diffraction methods. The crystal system is mono-clinic, space group is P2₁/c, Z = 4, a = 1.3741(1) nm, b = 2.6737(1) nm, c = 1.6690(1) nm, β= 101.278(1)°. The steric congestion between the neighbouring butoxyl groups causes the distortion of the ring core of phthalocyanine (Pc) into a saddle shape conformation. In the crystal structure, molecules stack along a axis forming one-dimensional packing structure and there are two molecular overlap types which appear in turn with different distances between molecules, overlap area and angle.     

The role of β18-β19 loop structure in insecticidal activity of Cry1Ac toxin from Bacillus thuringiensis

The β18-β19 loop in domain Ⅲ of Cry1Ac toxin is unique among Bacillus thuringlensis Cry proteins. In this study, the role of the loop structure in insecticidal activity of Cry1Ac toxin was investigated. Alanine scanning mutations within the loop were initially generated and most mutants were over-expressed and reduced toxicity at different degrees, except mutant N546A that showed almost 2 times enhanced toxicity against Helicoverpa armigera larvae. Further mutagenic analysis of N546 revealed that a charged amino acid in this position would cause very unfavorable influence on insecticidal activity. In addition, the deletion of N546 led to protein instability because of destruction of the loop integrity. Besides, mutant W544F was much more toxic than W544Y, indicating that hydrophobic nature of the position was important for maintaining the stability and activity of Cry1Ac protein. These findings are the first biological evidence for a structural function of β18-β19 loop in insecticidal activity of the Cry1Ac toxin.     

The role of β18-β19 loop structure in insecticidal activity of Cry1Ac toxin from Bacillus thuringiensis

The β18-β19 loop in domain III of Cry1Ac toxin is unique among Bacillus thuringiensis Cry proteins. In this study, the role of the loop structure in insecticidal activity of Cry1Ac toxin was investigated. Alanine scanning mutations within the loop were initially generated and most mutants were over-expressed and reduced toxicity at different degrees, except mutant N546A that showed almost 2 times enhanced toxicity against Helicoverpa armigera larvae. Further mutagenic analysis of N546 revealed that a charged amino acid in this position would cause very unfavorable influence on insecticidal activity. In addition, the deletion of N546 led to protein instability because of destruction of the loop integrity. Besides, mutant W544F was much more toxic than W544Y, indicating that hydrophobic nature of the position was important for maintaining the stability and activity of Cry1Ac protein. These findings are the first biological evidence for a structural function of β18-β19 loop in insecticidal activity of the Cry1Ac toxin.     

Structure of the human κ-opioid receptor in complex with JDTic

Opioid receptors mediate the actions of endogenous and exogenous opioids on many physiological processes, including the regulation of pain, respiratory drive, mood, and--in the case of κ-opioid receptor (κ-OR)--dysphoria and psychotomimesis. Here we report the crystal structure of the human κ-OR in complex with the selective antagonist JDTic, arranged in parallel dimers, at 2.9?? resolution. The structure reveals important features of the ligand-binding pocket that contribute to the high affinity and subtype selectivity of JDTic for the human κ-OR. Modelling of other important κ-OR-selective ligands, including the morphinan-derived antagonists norbinaltorphimine and 5'-guanidinonaltrindole, and the diterpene agonist salvinorin A analogue RB-64, reveals both common and distinct features for binding these diverse chemotypes. Analysis of site-directed mutagenesis and ligand structure-activity relationships confirms the interactions observed in the crystal structure, thereby providing a molecular explanation for κ-OR subtype selectivity, and essential insights for the design of compounds with new pharmacological properties targeting the human κ-OR.     

Isolation of tomato proteinase inhibitor Ⅱ gene and the function of its intron

The genomic DNA sequence of tomato proteinase inhibitor Ⅱ gene (named tin2i, whose accession number in GenBank is AF007240) was isolated by PCR techniques. The intron sequence (TPI), with a length of 109 bp, owns typical structures of GT/AG dinucleotides at both ends and high content of AT base pairs which accounts for 80.7% of the total nucleotides. As shown by recombination experiment, the TPI sequence could efficiently promote the expression of the reporter gene gusA and this effect was independent of the position and orientation of the intron, thus showing its role as an enhancer. Such experiments as gel retardation assays, GUS histochemical staining and GUS fluorometric assays further demonstrated that TPI sequence maybe has promoter-like activity.     

Selectivity of Crystal Growth Direction in Layered Double Hydroxides

Investigation of selectivity of crystal growth direction in layered double hydroxides is helpful to control their particle sizes in different directions. Mg-AI layered double hydroxides (LDHs) were synthesized using a coprecipitation method. The influences of aging temperature, aging time, and Mg/AI molar ratio on the crystal structure, the LDHs particle size, and the selectivity of crystal growth in different directions wereinvestigated. The results show that the size of the crystallites in the a direction is larger than that in the c direction for all experimental conditions, indicating faster crystal growth in the a direction than in the c direction. The crystallite sizes in the a and c directions both increase with decreasing Mg/AI molar ratio but with less difference between the sizes in the two directions. Therefore, the crystal growth rate in the c direction increases more than that in the a direction as the Mg/AI molar ratio decreases. The influence of the aging time, aging temperature, and Mg/AI molar ratio on the selectivity of the crystal growth direction can be used to prepare LDHs with selected sizes in the a and c directions.