The External TEA Binding Site and C-Type Inactivation in Voltage-Gated Potassium Channels

AbstractThe location of the tetraethylammonium (TEA) binding site in the outer vestibule of K+ channels, and the mechanism by which external TEA slows C-type inactivation, have been considered well-understood. The prevailing model has been that TEA is coordinated by four amino acid side chains at the position equivalent to Shaker T449, and that TEA prevents a constriction that underlies inactivation via a foot-in-the-door mechanism at this same position. However, a growing body of evidence has suggested that this picture may not be entirely correct. In this study, we reexamined these two issues, using both the Kv2.1 and Shaker potassium channels. In contrast to results previously obtained with Shaker, substitution of the tyrosine at Kv2.1 position 380 (equivalent to Shaker 449) with a threonine or cysteine had a relatively minor effect on TEA potency. In both Kv2.1 and Shaker, modification of cysteines at position 380/449 by 2-(trimethylammonium)ethyl methanethiosulfonate (MTSET) proceeded at identical rates in the absence and presence of TEA. Additional experiments in Shaker demonstrated that TEA bound well to C-type inactivated channels, but did not interfere with MTSET modification of C449 in inactivated channels. Together, these findings rule out the possibility that TEA binding involves an intimate interaction with the four side chains at the position equivalent to Shaker 449. Moreover, these results argue against the model whereby TEA slows inactivation via a foot-in-the-door mechanism at position 449, and also argue against the hypothesis that the position 449 side chains move toward the center of the conduction pathway during inactivation. Occupancy by TEA completely prevented MTSET modification of a cysteine in the outer-vestibule turret (Kv2.1 position 356/Shaker position 425), which has been shown to interfere with both TEA binding and the interaction of K+ with an external binding site. Together, these data suggest that TEA is stabilized in a more external position in the outer vestibule, and does not bind via direct coordination with any specific outer-vestibule residues

Control of Single Channel Conductance in the Outer Vestibule of the Kv2.1 Potassium Channel

Current magnitude in Kv2.1 potassium channels is modulated by external [K+]. In contrast to behavior expected from the change in electrochemical driving force, outward current through Kv2.1 channels becomes larger when extracellular [K+] is increased within the physiological range. The mechanism that underlies this unusual property involves the opening of Kv2.1 channels into one of two different outer vestibule conformations, which are defined by their sensitivity to TEA. Channels that open into a TEA-sensitive conformation generate larger macroscopic currents, whereas channels that open into a TEA-insensitive conformation generate smaller macroscopic currents. At higher [K+], more channels open into the TEA-sensitive conformation. In this manuscript, we examined the mechanism by which the conformational change produced a change in current magnitude. We started by testing the simplest hypothesis: that each pharmacologically defined channel conformation produces a different single channel conductance, one smaller and one larger, and that the [K+]-dependent change in current magnitude reflects the [K+]-dependent change in the percentage of channels that open into each of the two conformations. Using single channel and macroscopic recordings, as well as hidden Markov modeling, we were able to quantitatively account for [K+]-dependent regulation of macroscopic current with this model. Combined with previously published work, these results support a model whereby an outer vestibule lysine interferes with K+ flux through the channel, and that the [K+]-dependent change in orientation of this lysine alters single channel conductance by changing the level of this interference. Moreover, these results provide an experimental example of single channel conductance being modulated at the outer end of the conduction pathway by a mechanism that involves channel activation into open states with different outer vestibule conformations

Scaling relation for determining the critical threshold for continuum percolation of overlapping discs of two sizes

We study continuum percolation of overlapping circular discs of two sizes. We propose a phenomenological scaling equation for the increase in the effective size of the larger discs due to the presence of the smaller discs. The critical percolation threshold as a function of the ratio of sizes of discs, for different values of the relative areal densities of two discs, can be described in terms of a scaling function of only one variable. The recent accurate Monte Carlo estimates of critical threshold by Quintanilla and Ziff [Phys. Rev. E, 76 051115 (2007)] are in very good agreement with the proposed scaling relation.Comment: 4 pages, 3 figure

Variational Quantum Eigensolver for SU(NN) Fermions

Variational quantum algorithms aim at harnessing the power of noisy intermediate-scale quantum computers, by using a classical optimizer to train a parameterized quantum circuit to solve tractable quantum problems. The variational quantum eigensolver is one of the aforementioned algorithms designed to determine the ground-state of many-body Hamiltonians. Here, we apply the variational quantum eigensolver to study the ground-state properties of $N$-component fermions. With such knowledge, we study the persistent current of interacting SU($N$) fermions, which is employed to reliably map out the different quantum phases of the system. Our approach lays out the basis for a current-based quantum simulator of many-body systems that can be implemented on noisy intermediate-scale quantum computers.Comment: 9 pages, 8 figure

Quantum critical point in a periodic Anderson model

We investigate the symmetric Periodic Anderson Model (PAM) on a three-dimensional cubic lattice with nearest-neighbor hopping and hybridization matrix elements. Using Gutzwiller's variational method and the Hubbard-III approximation (which corresponds to the exact solution of an appropriate Falicov-Kimball model in infinite dimensions) we demonstrate the existence of a quantum critical point at zero temperature. Below a critical value $V_c$ of the hybridization (or above a critical interaction $U_c$) the system is an {\em insulator} in Gutzwiller's and a {\em semi-metal} in Hubbard's approach, whereas above $V_c$ (below $U_c$) it behaves like a metal in both approximations. These predictions are compared with the density of states of the $d$- and $f$-bands calculated from Quantum Monte Carlo and NRG calculations. Our conclusion is that the half-filled symmetric PAM contains a {\em metal-semimetal transition}, not a metal-insulator transition as has been suggested previously.Comment: ReVteX, 10 pages, 2 EPS figures. Minor corrections made in the text and in the figure captions from the first version. More references added. Accepted for publication in Physical Review

The periodic Anderson model from the atomic limit and FeSi

The exact Green's functions of the periodic Anderson model for $U\to \infty$ are formally expressed within the cumulant expansion in terms of an effective cumulant. Here we resort to a calculation in which this quantity is approximated by the value it takes for the exactly soluble atomic limit of the same model. In the Kondo region a spectral density is obtained that shows near the Fermi surface a structure with the properties of the Kondo peak. Approximate expressions are obtained for the static conductivity $$ and magnetic susceptibility $\chi (T)$ of the PAM, and they are employed to fit the experimental values of FeSi, a compound that behaves like a Kondo insulator with both quantities vanishing rapidly for $T\to 0$. Assuming that the system is in the intermediate valence region, it was possible to find good agreement between theory and experiment for these two properties by employing the same set of parameters. It is shown that in the present model the hybridization is responsible for the relaxation mechanism of the conduction electrons.Comment: 26 pages and 8 figure

The cosmic-ray electron flux measured by the PAMELA experiment between 1 and 625 GeV

Precision measurements of the electron component in the cosmic radiation provide important information about the origin and propagation of cosmic rays in the Galaxy. Here we present new results regarding negatively charged electrons between 1 and 625 GeV performed by the satellite-borne experiment PAMELA. This is the first time that cosmic-ray electrons have been identified above 50 GeV. The electron spectrum can be described with a single power law energy dependence with spectral index -3.18 +- 0.05 above the energy region influenced by the solar wind (> 30 GeV). No significant spectral features are observed and the data can be interpreted in terms of conventional diffusive propagation models. However, the data are also consistent with models including new cosmic-ray sources that could explain the rise in the positron fraction.Comment: 11 pages, 3 figures, accepted for publication in PR

Electron/pion separation with an Emulsion Cloud Chamber by using a Neural Network

We have studied the performance of a new algorithm for electron/pion separation in an Emulsion Cloud Chamber (ECC) made of lead and nuclear emulsion films. The software for separation consists of two parts: a shower reconstruction algorithm and a Neural Network that assigns to each reconstructed shower the probability to be an electron or a pion. The performance has been studied for the ECC of the OPERA experiment [1]. The $e/\pi$ separation algorithm has been optimized by using a detailed Monte Carlo simulation of the ECC and tested on real data taken at CERN (pion beams) and at DESY (electron beams). The algorithm allows to achieve a 90% electron identification efficiency with a pion misidentification smaller than 1% for energies higher than 2 GeV

PAMELA Measurements of Cosmic-ray Proton and Helium Spectra

Protons and helium nuclei are the most abundant components of the cosmic radiation. Precise measurements of their fluxes are needed to understand the acceleration and subsequent propagation of cosmic rays in the Galaxy. We report precision measurements of the proton and helium spectra in the rigidity range 1 GV-1.2 TV performed by the satellite-borne experiment PAMELA. We find that the spectral shapes of these two species are different and cannot be well described by a single power law. These data challenge the current paradigm of cosmic-ray acceleration in supernova remnants followed by diffusive propagation in the Galaxy. More complex processes of acceleration and propagation of cosmic rays are required to explain the spectral structures observed in our data.Comment: 13 pages, 4 figures, link to SOM (with tables) in the references. This manuscript has been accepted for publication in Science. This version has not undergone final editing. Please refer to the complete version of record at http://www.sciencemag.org/ [www.sciencemag.org

Large Animal Models in Regenerative Medicine and Tissue Engineering: To Do or Not to Do

Rapid developments in Regenerative Medicine and Tissue Engineering has witnessed an increasing drive toward clinical translation of breakthrough technologies. However, the progression of promising preclinical data to achieve successful clinical market authorisation remains a bottleneck. One hurdle for progress to the clinic is the transition from small animal research to advanced preclinical studies in large animals to test safety and efficacy of products. Notwithstanding this, to draw meaningful and reliable conclusions from animal experiments it is critical that the species and disease model of choice is relevant to answer the research question as well as the clinical problem. Selecting the most appropriate animal model requires in-depth knowledge of specific species and breeds to ascertain the adequacy of the model and outcome measures that closely mirror the clinical situation. Traditional reductionist approaches in animal experiments, which often do not sufficiently reflect the studied disease, are still the norm and can result in a disconnect in outcomes observed between animal studies and clinical trials. To address these concerns a reconsideration in approach will be required. This should include a stepwise approach using in vitro and ex vivo experiments as well as in silico modeling to minimize the need for in vivo studies for screening and early development studies, followed by large animal models which more closely resemble human disease. Naturally occurring, or spontaneous diseases in large animals remain a largely untapped resource, and given the similarities in pathophysiology to humans they not only allow for studying new treatment strategies but also disease etiology and prevention. Naturally occurring disease models, particularly for longer lived large animal species, allow for studying disorders at an age when the disease is most prevalent. As these diseases are usually also a concern in the chosen veterinary species they would be beneficiaries of newly developed therapies. Improved awareness of the progress in animal models is mutually beneficial for animals, researchers, human and veterinary patients. In this overview we describe advantages and disadvantages of various animal models including domesticated and companion animals used in regenerative medicine and tissue engineering to provide an informed choice of disease-relevant animal models