Visualization of class A GPCR oligomerization by image-based fluorescence fluctuation spectroscopy

G protein-coupled receptors (GPCRs) represent the largest class of cell surface receptors conveying extracellular information into intracellular signals. Many GPCRs have been shown to be able to oligomerize and it is firmly established that Class C GPCRs (e.g. metabotropic glutamate receptors) function as obligate dimers. However, the oligomerization capability of the larger Class A GPCRs (e.g. comprising the β-adrenergic receptors (β-ARs)) is still, despite decades of research, highly debated. Here we assess the oligomerization behavior of three prototypical Class A GPCRs, the β1-ARs, β2-ARs, and muscarinic M2Rs in single, intact cells. We combine two image correlation spectroscopy methods based on molecular brightness, i.e. the analysis of fluorescence fluctuations over space and over time, and thereby provide an assay able to robustly and precisely quantify the degree of oligomerization of GPCRs. In addition, we provide a comparison between two labelling strategies, namely C-terminally-attached fluorescent proteins and N-terminally-attached SNAP-tags, in order to rule out effects arising from potential fluorescent protein-driven oligomerization. The degree of GPCR oligomerization is expressed with respect to a set of previously reported as well as newly established monomeric or dimeric control constructs. Our data reveal that all three prototypical GPRCs studied display, under unstimulated conditions, a prevalently monomeric fingerprint. Only the β2-AR shows a slight degree of oligomerization. From a methodological point of view, our study suggests three key aspects. First, the combination of two image correlation spectroscopy methods allows addressing cells transiently expressing high concentrations of membrane receptors, far from the single molecule regime, at a density where the kinetic equilibrium should favor dimers and higher-order oligomers. Second, our methodological approach, allows to selectively target cell membrane regions devoid of artificial oligomerization hot-spots (such as vesicles). Third, our data suggest that the β1-AR appears to be a superior monomeric control than the widely used membrane protein CD86. Taken together, we suggest that our combined image correlation spectroscopy method is a powerful approach to assess the oligomerization behavior of GPCRs in intact cells at high expression levels

Fire and plant evolution

3 páginas, 1 figura.Peer reviewe

Open Heterotic Strings

We classify potential cosmic strings according to the topological charge measurable outside the string core. We conjecture that in string theory it is this charge that governs the stability of long strings. This would imply that the SO(32) heterotic string can have endpoints, but not the E_8 x E_8 heterotic string. We give various arguments in support of this conclusion.Comment: 15 pages. v.2: typos, references correcte

Quantum impurity solvers using a slave rotor representation

We introduce a representation of electron operators as a product of a spin-carry ing fermion and of a phase variable dual to the total charge (slave quantum rotor). Based on this representation, a new method is proposed for solving multi-orbital Anderson quantum impurity models at finite interaction strength U. It consists in a set of coupled integral equations for the auxiliary field Green's functions, which can be derived from a controlled saddle-point in the limit of a large number of field components. In contrast to some finite-U extensions of the non-crossing approximation, the new method provides a smooth interpolation between the atomic limit and the weak-coupling limit, and does not display violation of causality at low-frequency. We demonstrate that this impurity solver can be applied in the context of Dynamical Mean-Field Theory, at or close to half-filling. Good agreement with established results on the Mott transition is found, and large values of the orbital degeneracy can be investigated at low computational cost.Comment: 18 pages, 15 figure

Charge and spin density wave ordering transitions in strongly correlated metals

We study the quantum transition from a strongly correlated metal, with heavy fermionic quasiparticles, to a metal with commensurate charge or spin density wave order. To this end, we introduce and numerically analyze a large dimensionality model of Ising spins in a transverse field, coupled to two species of fermions; the analysis borrows heavily from recent progress in the solution of the Hubbard model in large dimensions. At low energies, the Ising order parameter fluctuations are characterized by the critical exponent $z \nu = 1$, while above an energy scale, $\Gamma$, there is a crossover to $z\nu = 1/2$ criticality. We show that $\Gamma$ is of the order of the width of the heavy quasiparticle band, and can be made arbitrarily small for a correlated metal close to a Mott-Hubbard insulator. Therefore, such a correlated metal has a significant intermediate energy range of $z\nu=1/2$ behavior, a single particle spectrum with a narrow quasiparticle band, and well-formed analogs of the lower and upper Hubbard bands; we suggest that these features are intimately related in general.Comment: 14 pages, REVTEX 3.0, 2 postscript figure

The Numerical Renormalization Group Method for correlated electrons

The Numerical Renormalization Group method (NRG) has been developed by Wilson in the 1970's to investigate the Kondo problem. The NRG allows the non-perturbative calculation of static and dynamic properties for a variety of impurity models. In addition, this method has been recently generalized to lattice models within the Dynamical Mean Field Theory. This paper gives a brief historical overview of the development of the NRG and discusses its application to the Hubbard model; in particular the results for the Mott metal-insulator transition at low temperatures.Comment: 14 pages, 7 eps-figures include

Two-site dynamical mean-field theory

It is shown that a minimum realization of the dynamical mean-field theory (DMFT) can be achieved by mapping a correlated lattice model onto an impurity model in which the impurity is coupled to an uncorrelated bath that consists of a single site only. The two-site impurity model can be solved exactly. The mapping is approximate. The self-consistency conditions are constructed in a way that the resulting ``two-site DMFT'' reduces to the previously discussed linearized DMFT for the Mott transition. It is demonstrated that a reasonable description of the mean-field physics is possible with a minimum computational effort. This qualifies the simple two-site DMFT for a systematic study of more complex lattice models which cannot be treated by the full DMFT in a feasible way. To show the strengths and limitations of the new approach, the single-band Hubbard model is investigated in detail. The predictions of the two-site DMFT are compared with results of the full DMFT. Internal consistency checks are performed which concern the Luttinger sum rule, other Fermi-liquid relations and thermodynamic consistency.Comment: LaTeX, 14 pages, 8 eps figures included, Phys. Rev. B (in press

Transport properties of strongly correlated metals:a dynamical mean-field approach

The temperature dependence of the transport properties of the metallic phase of a frustrated Hubbard model on the hypercubic lattice at half-filling are calculated. Dynamical mean-field theory, which maps the Hubbard model onto a single impurity Anderson model that is solved self-consistently, and becomes exact in the limit of large dimensionality, is used. As the temperature increases there is a smooth crossover from coherent Fermi liquid excitations at low temperatures to incoherent excitations at high temperatures. This crossover leads to a non-monotonic temperature dependence for the resistance, thermopower, and Hall coefficient, unlike in conventional metals. The resistance smoothly increases from a quadratic temperature dependence at low temperatures to large values which can exceed the Mott-Ioffe-Regel value, hbar a/e^2 (where "a" is a lattice constant) associated with mean-free paths less than a lattice constant. Further signatures of the thermal destruction of quasiparticle excitations are a peak in the thermopower and the absence of a Drude peak in the optical conductivity. The results presented here are relevant to a wide range of strongly correlated metals, including transition metal oxides, strontium ruthenates, and organic metals.Comment: 19 pages, 9 eps figure

Search for direct production of charginos and neutralinos in events with three leptons and missing transverse momentum in √s = 7 TeV pp collisions with the ATLAS detector

A search for the direct production of charginos and neutralinos in final states with three electrons or muons and missing transverse momentum is presented. The analysis is based on 4.7 fb−1 of proton–proton collision data delivered by the Large Hadron Collider and recorded with the ATLAS detector. Observations are consistent with Standard Model expectations in three signal regions that are either depleted or enriched in Z-boson decays. Upper limits at 95% confidence level are set in R-parity conserving phenomenological minimal supersymmetric models and in simplified models, significantly extending previous results

Jet size dependence of single jet suppression in lead-lead collisions at sqrt(s(NN)) = 2.76 TeV with the ATLAS detector at the LHC

Measurements of inclusive jet suppression in heavy ion collisions at the LHC provide direct sensitivity to the physics of jet quenching. In a sample of lead-lead collisions at sqrt(s) = 2.76 TeV corresponding to an integrated luminosity of approximately 7 inverse microbarns, ATLAS has measured jets with a calorimeter over the pseudorapidity interval |eta| < 2.1 and over the transverse momentum range 38 < pT < 210 GeV. Jets were reconstructed using the anti-kt algorithm with values for the distance parameter that determines the nominal jet radius of R = 0.2, 0.3, 0.4 and 0.5. The centrality dependence of the jet yield is characterized by the jet "central-to-peripheral ratio," Rcp. Jet production is found to be suppressed by approximately a factor of two in the 10% most central collisions relative to peripheral collisions. Rcp varies smoothly with centrality as characterized by the number of participating nucleons. The observed suppression is only weakly dependent on jet radius and transverse momentum. These results provide the first direct measurement of inclusive jet suppression in heavy ion collisions and complement previous measurements of dijet transverse energy imbalance at the LHC.Comment: 15 pages plus author list (30 pages total), 8 figures, 2 tables, submitted to Physics Letters B. All figures including auxiliary figures are available at http://atlas.web.cern.ch/Atlas/GROUPS/PHYSICS/PAPERS/HION-2011-02