Structural Basis for the Function of the C-Terminal Proton Release Pathway in the Calcium Pump

The calcium pump (sarco/endoplasmic reticulum Ca²⁺-ATPase, SERCA) plays a major role in calcium homeostasis in muscle cells by clearing cytosolic Ca²⁺ during muscle relaxation. Active Ca²⁺ transport by SERCA involves the structural transition from a low-Ca²⁺ affinity E2 state toward a high-Ca²⁺ affinity E1 state of the pump. This structural transition is accompanied by the countertransport of protons to stabilize the negative charge and maintain the structural integrity of the transport sites and partially compensate for the positive charges of the two Ca²⁺ ions passing through the membrane. X-ray crystallography studies have suggested that a hydrated pore located at the C-terminal domain of SERCA serves as a conduit for proton countertransport, but the existence and function of this pathway have not yet been fully characterized. We used atomistic simulations to demonstrate that in the protonated E2 state and the absence of initially bound water molecules, the C-terminal pore becomes hydrated in the nanosecond timescale. Hydration of the C-terminal pore is accompanied by the formation of water wires that connect the transport sites with the cytosol. Water wires are known as ubiquitous proton-transport devices in biological systems, thus supporting the notion that the C-terminal domain serves as a conduit for proton release. Additional simulations showed that the release of a single proton from the transport sites induces bending of transmembrane helix M5 and the interaction between residues Arg762 and Ser915. These structural changes create a physical barrier against full hydration of the pore and prevent the formation of hydrogen-bonded water wires once proton transport has occurred through this pore. Together, these findings support the notion that the C-terminal proton release pathway is a functional element of SERCA and also provide a mechanistic model for its operation in the catalytic cycle of the pump.     

Hydrogen from wet air and sunlight in a tandem photoelectrochemical cell

A solid-state photoelectrochemical (SSPEC) cell is an attractive approach for solar water splitting, especially when it comes to monolithic device design. In a SSPEC cell the electrodes distance is minimized, while the use of polymer-based membranes alleviates the need for liquid electrolytes, and at the same time they can separate the anode from the cathode. In this work, we have made and tested, firstly, a SSPEC cell with a Pt/C electrocatalyst as the cathode electrode, under purely gaseous conditions. The anode was supplied with air of 80% relative humidity (RH) and the cathode with argon. Secondly, we replaced the Pt/C cathode with a photocathode consisting of 2D photocatalytic g-C₃N₄, which was placed in tandem with the photoanode (tandem-SSPEC). The tandem configuration showed a three-fold enhancement in the obtained photovoltage and a steady-state photocurrent density. The mechanism of operation is discussed in view of recent advances in surface proton conduction in absorbed water layers. The presented SSPEC cell is based on earth-abundant materials and provides a way towards systems of artificial photosynthesis, especially for areas where water sources are scarce and electrical grid infrastructure is limited or nonexistent. The only requirements to make hydrogen are humidity and sunlight.     

Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

An Acoustic Converter Used to Measure Ionizing Radiation Energy Loss

A monitor is proposed based on ultrasonic production when ionizing radiation passes through a medium. The recording element is a 0.2 mm aluminum plate mounted in a ceramic acoustic converter AC in the form of a wedge of thickness 2 mm. The low plate thickness minimizes the beam parameter distortion, while special technology used in the AC provides high sensitivity. The device has been calibrated in the proton beam from the ITEP accelerator at 200 MeV with 2·10⁹–6·10¹⁰ particles in a pulse and a pulse length of 70 nsec.     

Preparation and characterization of proton‐exchange hybrid membranes

Two types of composite were prepared, based on a thermoplastic polymer, polyvinylidene fluoride (PVDF), and an elastomer, ethylene‐propylene‐diene terpolymer (EPDM), respectively. We obtained both series by addition of an inorganic proton‐conducting antimonic acid derivative (HSb) and polystyrene crosslinked with a small percentage of divinylbenzene (PS‐co‐DVB). From these composites, membranes were obtained and subjected to a heterogeneous‐phase sulfonation reaction with chlorosulfonic acid. All experimental materials were characterized from a morphological and electrical point of view, by means of techniques such as differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), non‐isothermal crystallization and complex impedance analysis. Copyright © 2005 Society of Chemical Industry     

Structural Description of Polyaromatic Nucleus in Residue

The proton nuclear magnetic resonance spectroscopy （1H-NMR）, the synchronous fluorescence spectrometry （SFS） and the ruthenium ions catalyzed oxidation （RICO） method were used to determine the chemical structure of polyaromatic nucleus in Oman residue fractions. The results of 1H-NMR analyses showed that the average numbers of aromatic rings in the aromatics, resins and asphaltenes units were 3.2, 5.6 and 8.2, respectively. SFS was used to investigate the distribution of aromatic rings in residue fractions, the main distribution range of aromatic rings in aromatics, resins and asphaltenes were 3 4 rings, 3--5 rings and more than 5 rings, respectively. The aromatic network in residue fractions was oxidized to produce numerous carboxylic acids. The types and content of benzenepolycarboxylic acids, such as phthalic acid, benzenetricarboxylic acids, benzenetetracarboxylic acids, benzenepentacarboxylic acid and benzenehexacarboxylic acid disclosed the condensed types of aromatic nuclei in the core. The biphenyl fraction （BIPH）, the cata-condensed fraction （CATA）, the peri-condensed fraction （PERI） and the condensed index （BCI） were calculated based on the benzenepolycarboxylic acids formed. The results implied that there was less biphenyl type structures in all residue fractions. The aromatics fraction was almost composed of the cata-condensed type system, and the asphaltenes fraction was wholly composed of the peri-condensed type system, while in the resins fraction co-existed the two types, herein the peri-con- densed type was predominant over the cata-condensed type. Based on the analytical results obtained in the study, the components --aromatics, resins and asphaltenes -- were given the likely structural models.     

质子和中子的单粒子效应等效性实验研究

通过实验方法确定了高能质子和中子引起的单粒子效应等效关系，实验中采用金箔散射法降低了质子束流强度，并利用热释光剂量计（TLD）进行质子注量的监测，采用新研制的存储器长线实时监测系统，进行了64K位至4M位的SRAM器件单粒子效应实验，确定了两种粒子引起单粒子效应等效关系。     

聚乙烯醇膜的阻醇及导电性能(Ⅱ)戊二醛交联聚乙烯醇膜

为提高直接甲醇燃料电池(DMFC)用膜的阻醇性能，采用在渗透蒸发领域广泛使用且具有良好分离效果的聚乙烯醇(PVA)为主要材料，制备了戊二醛(GA)交联PVA膜，考察了此类膜的阻醇及质子导电能力。PVA(GA)膜的阻醇效果较目前在DMFC中广泛使用的Nation膜有显著提高，但其自身不具有质子导电能力，需外加电解质溶液以提高其电导率。     

基于多孔硅及MEMS技术的微型质子交换膜燃料电池

分析了研究和发展微型质子交换膜燃料电池(μPEMFC)的重要意义和应用前景,介绍了基于多孔硅及MEMS技术的μPEMFC最新研究动态,讨论了μPEMFC的结构设计、加工过程及性能,提出了μPEMFC仍存在的问题及其技术发展对策。     

Ammonium Accumulation Caused by Reduced Tonoplast V-ATPase Activity in Arabidopsis thaliana

Plant vacuoles are unique compartments that play a critical role in plant growth and development. The vacuolar H⁺-ATPase (V-ATPase), together with the vacuolar H⁺-pyrophosphatase (V-PPase), generates the proton motive force that regulates multiple cell functions and impacts all aspects of plant life. We investigated the effect of V-ATPase activity in the vacuole on plant growth and development. We used an Arabidopsis thaliana (L.) Heynh. double mutant, vha-a2 vha-a3, which lacks two tonoplast-localized isoforms of the membrane-integral V-ATPase subunit VHA-a. The mutant is viable but exhibits impaired growth and leaf chlorosis. Nitrate assimilation led to excessive ammonium accumulation in the shoot and lower nitrogen uptake, which exacerbated growth retardation of vha-a2 vha-a3. Ion homeostasis was disturbed in plants with missing VHA-a2 and VHA-a3 genes, which might be related to limited growth. The reduced growth and excessive ammonium accumulation of the double mutant was alleviated by potassium supplementation. Our results demonstrate that plants lacking the two tonoplast-localized subunits of V-ATPase can be viable, although with defective growth caused by multiple factors, which can be alleviated by adding potassium. This study provided a new insight into the relationship between V-ATPase, growth, and ammonium accumulation, and revealed the role of potassium in mitigating ammonium toxicity.