Thickness dependent magnetic anisotropy of ultrathin LCMO epitaxial thin films

The magnetic properties of La0.7Ca0.3MnO3 (LCMO) manganite thin films were studied with magnetometry and ferromagnetic resonance as a function of film thickness. They maintain the colossal magnetoresistance behavior with a pronounced metal-insulator transition around 150-200 K, except for the very thinnest films studied (3 nm). Nevertheless, LCMO films as thin as 3 nm remain ferromagnetic, without a decrease in saturation magnetization, indicating an absence of dead-layers, although below approx. 6 nm the films remain insulating at low temperature. Magnetization hysteresis loops reveal that the magnetic easy axes lie in the plane of the film for thicknesses in the range of 4-15 nm. Ferromagnetic resonance studies confirm that the easy axes are in-plane, and find a biaxial symmetry in-plane with two, perpendicular easy axes. The directions of the easy axes with respect to the crystallographic directions of the cubic SrTiO3 substrate differ by 45 degrees in 4 nm and 15 nm thick LCMO films.Comment: Presented at Intermag conference (Madrid, 2008). Accepted for publication in IEEE Transactions on Magnetic

Peptide Inhibitors of Kv1.5: An Option for the Treatment of Atrial Fibrillation

Abstract: The human voltage gated potassium channel Kv1.5 that conducts the IKurcurrent is akey determinant of the atrial action potential. Its mutations have been linked to hereditary formsof atrial fibrillation (AF), and the channel is an attractive target for the management of AF. Thedevelopment of IKurblockers to treat AF resulted in small molecule Kv1.5 inhibitors. The selectivityof the blocker for the target channel plays an important role in the potential therapeutic applicationof the drug candidate: the higher the selectivity, the lower the risk of side effects. In this respect,small molecule inhibitors of Kv1.5 are compromised due to their limited selectivity. A wide range ofpeptide toxins from venomous animals are targeting ion channels, including mammalian channels.These peptides usually have a much larger interacting surface with the ion channel compared tosmall molecule inhibitors and thus, generally confer higher selectivity to the peptide blockers. Wefound two peptides in the literature, which inhibited IKur: Ts6 and Osu1. Their affinity and selectivityfor Kv1.5 can be improved by rational drug design in which their amino acid sequences could bemodified in a targeted way guided by in silico docking experiments

Elucidation of the binding mode of organic polysulfides on the human TRPA1 receptor

Introduction: Previous studies have established that endogenous inorganic polysulfides have significant biological actions activating the Transient Receptor Potential Ankyrin 1 (TRPA1) receptor. Organic polysulfides exert similar effects, but they are much more stable molecules, therefore these compounds are more suitable as drugs. In this study, we aimed to better understand the mechanism of action of organic polysulfides by identification of their binding site on the TRPA1 receptor.Methods: Polysulfides can readily interact with the thiol side chain of the cysteine residues of the protein. To investigate their role in the TRPA1 activation, we replaced several cysteine residues by alanine via site-directed mutagenesis. We searched for TRPA1 mutant variants with decreased or lost activating effect of the polysulfides, but with other functions remaining intact (such as the effects of non-electrophilic agonists and antagonists). The binding properties of the mutant receptors were analyzed by in silico molecular docking. Functional changes were tested by in vitro methods: calcium sensitive fluorescent flow cytometry, whole-cell patch-clamp and radioactive calcium-45 liquid scintillation counting.Results: The cysteines forming the conventional binding site of electrophilic agonists, namely C621, C641 and C665 also bind the organic polysulfides, with the key role of C621. However, only their combined mutation abolished completely the organic polysulfide-induced activation of the receptor.Discussion: Since previous papers provided evidence that organic polysulfides exert analgesic and anti-inflammatory actions in different in vivo animal models, we anticipate that the development of TRPA1-targeted, organic polysulfide-based drugs will be promoted by this identification of the binding site

Multiple mechanisms contribute to fluorometry signals from the voltage-gated proton channel

Abstract Voltage-clamp fluorometry (VCF) supplies information about the conformational changes of voltage-gated proteins. Changes in the fluorescence intensity of the dye attached to a part of the protein that undergoes a conformational rearrangement upon the alteration of the membrane potential by electrodes constitute the signal. The VCF signal is generated by quenching and dequenching of the fluorescence as the dye traverses various local environments. Here we studied the VCF signal generation, using the Hv1 voltage-gated proton channel as a tool, which shares a similar voltage-sensor structure with voltage-gated ion channels but lacks an ion-conducting pore. Using mutagenesis and lipids added to the extracellular solution we found that the signal is generated by the combined effects of lipids during movement of the dye relative to the plane of the membrane and by quenching amino acids. Our 3-state model recapitulates the VCF signals of the various mutants and is compatible with the accepted model of two major voltage-sensor movements

Functional Voltage-Gated Sodium Channels Are Present in the Human B Cell Membrane

B cells express various ion channels, but the presence of voltage-gated sodium (NaV) channels has not been confirmed in the plasma membrane yet. In this study, we have identified several NaV channels, which are expressed in the human B cell membrane, by electrophysiological and molecular biology methods. The sensitivity of the detected sodium current to tetrodotoxin was between the values published for TTX-sensitive and TTX-insensitive channels, which suggests the co-existence of multiple NaV1 subtypes in the B cell membrane. This was confirmed by RT-qPCR results, which showed high expression of TTX-sensitive channels along with the lower expression of TTX-insensitive NaV1 channels. The biophysical characteristics of the currents also supported the expression of multiple NaV channels. In addition, we investigated the potential functional role of NaV channels by membrane potential measurements. Removal of Na+ from the extracellular solution caused a reversible hyperpolarization, supporting the role of NaV channels in shaping and maintaining the resting membrane potential. As this study was mainly limited to electrophysiological properties, we cannot exclude the possible non-canonical functions of these channels. This work concludes that the presence of voltage-gated sodium channels in the plasma membrane of human B cells should be recognized and accounted for in the future

Functional Voltage-Gated Sodium Channels Are Present in the Human B Cell Membrane

B cells express various ion channels, but the presence of voltage-gated sodium (NaV) channels has not been confirmed in the plasma membrane yet. In this study, we have identified several NaV channels, which are expressed in the human B cell membrane, by electrophysiological and molecular biology methods. The sensitivity of the detected sodium current to tetrodotoxin was between the values published for TTX-sensitive and TTX-insensitive channels, which suggests the co-existence of multiple NaV1 subtypes in the B cell membrane. This was confirmed by RT-qPCR results, which showed high expression of TTX-sensitive channels along with the lower expression of TTX-insensitive NaV1 channels. The biophysical characteristics of the currents also supported the expression of multiple NaV channels. In addition, we investigated the potential functional role of NaV channels by membrane potential measurements. Removal of Na+ from the extracellular solution caused a reversible hyperpolarization, supporting the role of NaV channels in shaping and maintaining the resting membrane potential. As this study was mainly limited to electrophysiological properties, we cannot exclude the possible non-canonical functions of these channels. This work concludes that the presence of voltage-gated sodium channels in the plasma membrane of human B cells should be recognized and accounted for in the future