Structure of an agonist-bound human A2A adenosine receptor

Activation of G protein-coupled receptors upon agonist binding is a critical step in the signaling cascade for this family of cell surface proteins. We report the crystal structure of the A(2A) adenosine receptor (A(2A)AR) bound to an agonist UK-432097 at 2.7 angstrom resolution. Relative to inactive, antagonist-bound A(2A)AR, the agonist-bound structure displays an outward tilt and rotation of the cytoplasmic half of helix VI, a movement of helix V, and an axial shift of helix III, resembling the changes associated with the active-state opsin structure. Additionally, a seesaw movement of helix VII and a shift of extracellular loop 3 are likely specific to A(2A)AR and its ligand. The results define the molecule UK-432097 as a "conformationally selective agonist" capable of receptor stabilization in a specific active-state configuration.     

Structure of the human dopamine D3 receptor in complex with a D2/D3 selective antagonist

Dopamine modulates movement, cognition, and emotion through activation of dopamine G protein-coupled receptors in the brain. The crystal structure of the human dopamine D3 receptor (D3R) in complex with the small molecule D2R/D3R-specific antagonist eticlopride reveals important features of the ligand binding pocket and extracellular loops. On the intracellular side of the receptor, a locked conformation of the ionic lock and two distinctly different conformations of intracellular loop 2 are observed. Docking of R-22, a D3R-selective antagonist, reveals an extracellular extension of the eticlopride binding site that comprises a second binding pocket for the aryl amide of R-22, which differs between the highly homologous D2R and D3R. This difference provides direction to the design of D3R-selective agents for treating drug abuse and other neuropsychiatric indications.     

Photoperiod in recirculation aquaculture systems and timing of seawater transfer affect seawater growth performance of Atlantic salmon (Salmo salar)

Production of Atlantic salmon smolts in recirculation aquaculture systems (RAS) is growing, and novel production protocols using continuous light in RAS are being implemented in the industry. In the present study, Atlantic Salmon parr were exposed to either a traditional protocol (short-day winter signal [12:12 L:D] for 6 weeks) or to continuous light. Both photoperiods were applied in freshwater (FW) and brackish water RAS. Salmon from all treatments were transferred to seawater pens at 200 and 600 g and grown until slaughter size. A control group was smoltified with a 6-week short-day winter signal and kept in FW until sea transfer at 100 g. Continuous light gave a higher growth rate in RAS but reduced feed intake and growth and increased feed conversion ratio during the first 8 weeks in seawater. However, at slaughter, fish exposed to continuous light was bigger than fish given a winter signal because of the higher growth rate in RAS. Slaughter weight was lowest in fish transferred to sea at 600 g, despite having the highest day-degree sum during their life span. The best performing group was the control group transferred at 100 g. All treatments handled transfer to seawater and survival and maturation were not affected by the treatments in RAS. The immune status was examined with a multigene expression assay on BioMark HD platform from parr stage to 5–7 months after seawater transfer. Overall, there was no significant effect of photoperiod or salinity on the expression of the selected immune genes. In sum, the results from this study indicate that using continuous light in RAS may have negative effects on performance shortly after transfer in fish transferred to sea at 200 g, whereas at 600 g, all treatments had reduced growth after transfer irrespective of treatment in RAS.     

Cytologic,histologic, microbiologic,and electron microscopic characterization of a canine Prototheca wickerhamii infection

An adult dog was presented for chronic cough and a recent development of ulcerated, erythematous nares with nasal discharge. Cytology of enlarged peripheral lymph nodes revealed many intracellular and extracellular organisms. These round or rarely oval organisms measured approximately 5-9 µm in diameter and frequently contained several globular structures, ranging from deeply basophilic to magenta. A thin, clear halo was present. Smaller 1-2 µm, magenta forms were also observed. Fungal culture yielded small, wet, raised, irregularly shaped, white to pale tan colonies. Microbiologic staining of cultured material revealed features suggestive of algae. Histopathology of the lymph nodes revealed marked granulomatous inflammation with intralesional algal organisms suggestive of Prototheca. Electron microscopic findings were also consistent with protothecosis. Polymerase chain reaction, followed by direct DNA sequencing, identified the organism as Prototheca wickerhamii. A brief literature review discussing protothecosis in veterinary medicine is included.     

Biased signaling pathways in β2-adrenergic receptor characterized by 19F-NMR

Extracellular ligand binding to G protein-coupled receptors (GPCRs) modulates G protein and β-arrestin signaling by changing the conformational states of the cytoplasmic region of the receptor. Using site-specific (19)F-NMR (fluorine-19 nuclear magnetic resonance) labels in the β(2)-adrenergic receptor (β(2)AR) in complexes with various ligands, we observed that the cytoplasmic ends of helices VI and VII adopt two major conformational states. Changes in the NMR signals reveal that agonist binding primarily shifts the equilibrium toward the G protein-specific active state of helix VI. In contrast, β-arrestin-biased ligands predominantly impact the conformational states of helix VII. The selective effects of different ligands on the conformational equilibria involving helices VI and VII provide insights into the long-range structural plasticity of β(2)AR in partial and biased agonist signaling.     

Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists

Chemokine receptors are critical regulators of cell migration in the context of immune surveillance, inflammation, and development. The G protein-coupled chemokine receptor CXCR4 is specifically implicated in cancer metastasis and HIV-1 infection. Here we report five independent crystal structures of CXCR4 bound to an antagonist small molecule IT1t and a cyclic peptide CVX15 at 2.5 to 3.2 angstrom resolution. All structures reveal a consistent homodimer with an interface including helices V and VI that may be involved in regulating signaling. The location and shape of the ligand-binding sites differ from other G protein-coupled receptors and are closer to the extracellular surface. These structures provide new clues about the interactions between CXCR4 and its natural ligand CXCL12, and with the HIV-1 glycoprotein gp120.     

Global circumnavigations: tracking year-round ranges of nonbreeding albatrosses

Although albatrosses are paradigms of oceanic specialization, their foraging areas and migration routes when not breeding remain essentially unknown. Our continuous remote tracking of 22 adult gray-headed albatrosses for over 30 bird-years reveals three distinct strategies: (i) Stay in breeding home range; (ii) make return migrations to a specific area of the southwest Indian Ocean; and (iii) make one or more global circumnavigations (the fastest in just 46 days). The consistencies in patterns, routes, and timings offer the first hope of identifying areas of critical habitat for nonbreeding albatrosses, wherein appropriate management of longline fisheries might alleviate the plight of the world's most threatened family of birds.     

Leben und WerkN. I. Vavilovs

Ohne Zusammenfassung     

Structural basis for allosteric regulation of GPCRs by sodium ions

Pharmacological responses of G protein-coupled receptors (GPCRs) can be fine-tuned by allosteric modulators. Structural studies of such effects have been limited due to the medium resolution of GPCR structures. We reengineered the human A(2A) adenosine receptor by replacing its third intracellular loop with apocytochrome b(562)RIL and solved the structure at 1.8 angstrom resolution. The high-resolution structure allowed us to identify 57 ordered water molecules inside the receptor comprising three major clusters. The central cluster harbors a putative sodium ion bound to the highly conserved aspartate residue Asp(2.50). Additionally, two cholesterols stabilize the conformation of helix VI, and one of 23 ordered lipids intercalates inside the ligand-binding pocket. These high-resolution details shed light on the potential role of structured water molecules, sodium ions, and lipids/cholesterol in GPCR stabilization and function.