Molecular identification of the CRAC channel by altered ion selectivity in a mutant of Orai

Recent RNA interference screens have identified several proteins that are essential for store-operated Ca2+ influx and Ca2+ release-activated Ca2+ (CRAC) channel activity in Drosophila and in mammals, including the transmembrane proteins Stim (stromal interaction molecule) and Orai. Stim probably functions as a sensor of luminal Ca2+ content and triggers activation of CRAC channels in the surface membrane after Ca2+ store depletion. Among three human homologues of Orai (also known as olf186-F), ORAI1 on chromosome 12 was found to be mutated in patients with severe combined immunodeficiency disease, and expression of wild-type Orai1 restored Ca2+ influx and CRAC channel activity in patient T cells. The overexpression of Stim and Orai together markedly increases CRAC current. However, it is not yet clear whether Stim or Orai actually forms the CRAC channel, or whether their expression simply limits CRAC channel activity mediated by a different channel-forming subunit. Here we show that interaction between wild-type Stim and Orai, assessed by co-immunoprecipitation, is greatly enhanced after treatment with thapsigargin to induce Ca2+ store depletion. By site-directed mutagenesis, we show that a point mutation from glutamate to aspartate at position 180 in the conserved S1-S2 loop of Orai transforms the ion selectivity properties of CRAC current from being Ca2+-selective with inward rectification to being selective for monovalent cations and outwardly rectifying. A charge-neutralizing mutation at the same position (glutamate to alanine) acts as a dominant-negative non-conducting subunit. Other charge-neutralizing mutants in the same loop express large inwardly rectifying CRAC current, and two of these exhibit reduced sensitivity to the channel blocker Gd3+. These results indicate that Orai itself forms the Ca2+-selectivity filter of the CRAC channel.     

Nano-Structured Carbide-Derived Carbon Films and Their Tribology

Carbide-derived carbon （CDC） is a form of carbon produced by reacting metal carbides, such as SiC or TiC, with halogens at temperatures high enough to produce fast kinetics, but too low to permit the rearrangement of the carbon atoms into an equilibrium graphitic structure. The structure of CDC is derivative of the original carbide structure and contains nanoscale porosity and both sp2 and sp3 bonded carbon in a variety of nanoscale structures. CDC can be produced as a thin film on hard carbides to improve their tribological performance. CDC coatings are distinguished by their low friction coefficients and high wear resistance in many important industrial environments and by their resistance to spallation and delamination. The tribology of CDC coatings on SiC surfaces is described in detail.     

The CRAC channel consists of a tetramer formed by Stim-induced dimerization of Orai dimers

Ca(2+)-release-activated Ca(2+) (CRAC) channels underlie sustained Ca(2+) signalling in lymphocytes and numerous other cells after Ca(2+) liberation from the endoplasmic reticulum (ER). RNA interference screening approaches identified two proteins, Stim and Orai, that together form the molecular basis for CRAC channel activity. Stim senses depletion of the ER Ca(2+) store and physically relays this information by translocating from the ER to junctions adjacent to the plasma membrane, and Orai embodies the pore of the plasma membrane calcium channel. A close interaction between Stim and Orai, identified by co-immunoprecipitation and by F?rster resonance energy transfer, is involved in the opening of the Ca(2+) channel formed by Orai subunits. Most ion channels are multimers of pore-forming subunits surrounding a central channel, which are preassembled in the ER and transported in their final stoichiometry to the plasma membrane. Here we show, by biochemical analysis after cross-linking in cell lysates and intact cells and by using non-denaturing gel electrophoresis without cross-linking, that Orai is predominantly a dimer in the plasma membrane under resting conditions. Moreover, single-molecule imaging of green fluorescent protein (GFP)-tagged Orai expressed in Xenopus oocytes showed predominantly two-step photobleaching, again consistent with a dimeric basal state. In contrast, co-expression of GFP-tagged Orai with the carboxy terminus of Stim as a cytosolic protein to activate the Orai channel without inducing Ca(2+) store depletion or clustering of Orai into punctae yielded mostly four-step photobleaching, consistent with a tetrameric stoichiometry of the active Orai channel. Interaction with the C terminus of Stim thus induces Orai dimers to dimerize, forming tetramers that constitute the Ca(2+)-selective pore. This represents a new mechanism in which assembly and activation of the functional ion channel are mediated by the same triggering molecule.     

Electronic nematicity above the structural and superconducting transition in BaFe2(As(1-x)P(x))2

Electronic nematicity, a unidirectional self-organized state that breaks the rotational symmetry of the underlying lattice, has been observed in the iron pnictide and copper oxide high-temperature superconductors. Whether nematicity plays an equally important role in these two systems is highly controversial. In iron pnictides, the nematicity has usually been associated with the tetragonal-to-orthorhombic structural transition at temperature T(s). Although recent experiments have provided hints of nematicity, they were performed either in the low-temperature orthorhombic phase or in the tetragonal phase under uniaxial strain, both of which break the 90° rotational C(4) symmetry. Therefore, the question remains open whether the nematicity can exist above T(s) without an external driving force. Here we report magnetic torque measurements of the isovalent-doping system BaFe(2)(As(1-x)P(x))(2), showing that the nematicity develops well above T(s) and, moreover, persists to the non-magnetic superconducting regime, resulting in a phase diagram similar to the pseudogap phase diagram of the copper oxides. By combining these results with synchrotron X-ray measurements, we identify two distinct temperatures-one at T*, signifying a true nematic transition, and the other at T(s) (相似文献   

Electrodeposition of Al from a 1-butylpyrrolidine-AlCl3 ionic liquid

The addition of 1-butylpyrrolidine to AlCl_3 results in the formation of an electrolyte that is suited to Al deposition.The feasibility of electrodepositing Al from the synthesized electrolyte was investigated.Several compositions of AlCl_3 and 1-butylpyrrolidine were prepared for this purpose.These mixtures show a different phase behavior at various compositions of AlCl_3 and 1-butylpyrrolidine.IR,Raman and NMR spectroscopy were employed to characterize the synthesized liquids.Among the prepared compositions,1:1.2 mol ratio of 1-butylpyrrolidine:AlCl_3 and the upper phase of 1:1.3 mol ratio of 1-butylpyrrolidine:AlCl_3 were found to be suitable for Al electrodeposition at room temperature(RT).Uniform and thick( mm thick) layers of Al were obtained on copper at RT.Al deposition occured from the cationic species of AlCl_3 xLty(where x r 2,y ? 1–2,and L ? 1-butylpyrrolidine) in this electrolyte.This behavior is contrary to the well investigated classic AlCl_3 based ionic liquids,where the deposition of Al occurs mainly from anionic Al2 Cl 7ions.