Structure and mechanism of mitochondrial proton-translocating transhydrogenase

Proton-translocating transhydrogenase (also known as nicotinamide nucleotide transhydrogenase (NNT)) is found in the plasma membranes of bacteria and the inner mitochondrial membranes of eukaryotes. NNT catalyses the transfer of a hydride between NADH and NADP+, coupled to the translocation of one proton across the membrane. Its main physiological function is the generation of NADPH, which is a substrate in anabolic reactions and a regulator of oxidative status; however, NNT may also fine-tune the Krebs cycle1,2. NNT deficiency causes familial glucocorticoid deficiency in humans and metabolic abnormalities in mice, similar to those observed in type II diabetes3,4. The catalytic mechanism of NNT has been proposed to involve a rotation of around 180° of the entire NADP(H)-binding domain that alternately participates in hydride transfer and proton-channel gating. However, owing to the lack of high-resolution structures of intact NNT, the details of this process remain unclear5,6. Here we present the cryo-electron microscopy structure of intact mammalian NNT in different conformational states. We show how the NADP(H)-binding domain opens the proton channel to the opposite sides of the membrane, and we provide structures of these two states. We also describe the catalytically important interfaces and linkers between the membrane and the soluble domains and their roles in nucleotide exchange. These structures enable us to propose a revised mechanism for a coupling process in NNT that is consistent with a large body of previous biochemical work. Our results are relevant to the development of currently unavailable NNT inhibitors, which may have therapeutic potential in ischaemia reperfusion injury, metabolic syndrome and some cancers7,8,9

The coupling mechanism of mammalian respiratory complex I

Mitochondrial complex I couples NADH:ubiquinone oxidoreduction to proton pumping by an unknown mechanism. Here, we present cryo-electron microscopy structures of ovine complex I in five different conditions, including turnover, at resolutions up to 2.3 to 2.5 angstroms. Resolved water molecules allowed us to experimentally define the proton translocation pathways. Quinone binds at three positions along the quinone cavity, as does the inhibitor rotenone that also binds within subunit ND4. Dramatic conformational changes around the quinone cavity couple the redox reaction to proton translocation during open-to-closed state transitions of the enzyme. In the induced deactive state, the open conformation is arrested by the ND6 subunit. We propose a detailed molecular coupling mechanism of complex I, which is an unexpected combination of conformational changes and electrostatic interactions

Cryo-EM grid optimization for membrane proteins

Cryo-EM grid preparation is an important bottleneck in protein structure determination, especially for membrane proteins, typically requiring screening of a large number of conditions. We systematically investigated the effects of buffer components, blotting conditions and grid types on the outcome of grid preparation of five different membrane protein samples. Aggregation was the most common type of problem which was addressed by changing detergents, salt concentration or reconstitution of proteins into nanodiscs or amphipols. We show that the optimal concentration of detergent is between 0.05 and 0.4% and that the presence of a low concentration of detergent with a high critical micellar concentration protects the proteins from denaturation at the air-water interface. Furthermore, we discuss the strategies for achieving an adequate ice thickness, particle coverage and orientation distribution on free ice and on support films. Our findings provide a clear roadmap for comprehensive screening of conditions for cryo-EM grid preparation of membrane proteins

Atomic quasi-Bragg diffraction in a magnetic field

We report on a new technique to split an atomic beam coherently with an easily adjustable splitting angle. In our experiment metastable helium atoms in the |{1s2s}^3S_1 M=1> state diffract from a polarization gradient light field formed by counterpropagating \sigma^+ and \sigma^- polarized laser beams in the presence of a homogeneous magnetic field. In the near-adiabatic regime, energy conservation allows the resonant exchange between magnetic energy and kinetic energy. As a consequence, symmetric diffraction of |M=0> or |M=-1> atoms in a single order is achieved, where the order can be chosen freely by tuning the magnetic field. We present experimental results up to 6th order diffraction (24 \hbar k momentum splitting, i.e., 2.21 m/s in transverse velocity) and present a simple theoretical model that stresses the similarity with conventional Bragg scattering. The resulting device constitutes a flexible, adjustable, large-angle, three-way coherent atomic beam splitter with many potential applications in atom optics and atom interferometry.Comment: 4 pages, 5 figure

Effects of macroscopic polarization in III-V nitride multi-quantum-wells

Huge built-in electric fields have been predicted to exist in wurtzite III-V nitrides thin films and multilayers. Such fields originate from heterointerface discontinuities of the macroscopic bulk polarization of the nitrides. Here we discuss the background theory, the role of spontaneous polarization in this context, and the practical implications of built-in polarization fields in nitride nanostructures. To support our arguments, we present detailed self-consistent tight-binding simulations of typical nitride QW structures in which polarization effects are dominant.Comment: 11 pages, 9 figures, uses revtex/epsf. submitted to PR

Closing the loop in exergaming - Health benefits of biocybernetic adaptation in senior adults

Exergames help senior players to get physically active by promoting fun and enjoyment while exercising. However, most exergames are not designed to produce recommended levels of exercise that elicit adequate physical responses for optimal training in the aged population. In this project, we developed physiological computing technologies to overcome this issue by making real-time adaptations in a custom exergame based on recommendations for targeted heart rate (HR) levels. This biocybernetic adaptation was evaluated against conventional cardiorespiratory training in a group of active senior adults through a floor-projected exergame and a smartwatch to record HR data. Results showed that the physiologically-augmented exergame leads players to exert around 40% more time in the recommended HR levels, compared to the conventional training, avoiding over exercising and maintaining good enjoyment levels. Finally, we made available our biocybernetic adaptation software tool to enable the creation of physiological adaptive videogames, permitting the replication of our study.info:eu-repo/semantics/publishedVersio

Strategies for Improving the Photocatalytic Hydrogen Evolution Reaction of Carbon Nitride-Based Catalysts

Published online:. OnlinePublDue to the depletion of fossil fuels and their-related environmental issues, sustainable, clean, and renewable energy is urgently needed to replace fossil fuel as the primary energy resource. Hydrogen is considered as one of the cleanest energies. Among the approaches to hydrogen production, photocatalysis is the most sustainable and renewable solar energy technique. Considering the low cost of fabrication, earth abundance, appropriate bandgap, and high performance, carbon nitride has attracted extensive attention as the catalyst for photocatalytic hydrogen production in the last two decades. In this review, the carbon nitride-based photocatalytic hydrogen production system, including the catalytic mechanism and the strategies for improving the photocatalytic performance is discussed. According to the photocatalytic processes, the strengthened mechanism of carbon nitride-based catalysts is particularly described in terms of boosting the excitation of electrons and holes, suppressing carriers recombination, and enhancing the utilization efficiency of photon-excited electron-hole. Finally, the current trends related to the screening design of superior photocatalytic hydrogen production systems are outlined, and the development direction of carbon nitride for hydrogen production is clarified.Xueze Chu, C.I. Sathish, Jae-Hun Yang, Xinwei Guan, Xiangwei Zhang, Liang Qiao, Kazunari Domen, Shaobin Wang, Ajayan Vinu, and Jiabao Y

Linking in situ charge accumulation to electronic structure in doped SrTiO3 reveals design principles for hydrogen-evolving photocatalysts

Recently, high solar-to-hydrogen efficiencies were demonstrated using La and Rh co-doped SrTiO3 (La,Rh:SrTiO3) incorporated into a low-cost and scalable Z-scheme device, known as a photocatalyst sheet. However, the unique properties that enable La,Rh:SrTiO3 to support this impressive performance are not fully understood. Combining in situ spectroelectrochemical measurements with density functional theory and photoelectron spectroscopy produces a depletion model of Rh:SrTiO3 and La,Rh:SrTiO3 photocatalyst sheets. This reveals remarkable properties, such as deep flatband potentials (+2 V versus the reversible hydrogen electrode) and a Rh oxidation state dependent reorganization of the electronic structure, involving the loss of a vacant Rh 4d mid-gap state. This reorganization enables Rh:SrTiO3 to be reduced by co-doping without compromising the p-type character. In situ time-resolved spectroscopies show that the electronic structure reorganization induced by Rh reduction controls the electron lifetime in photocatalyst sheets. In Rh:SrTiO3, enhanced lifetimes can only be obtained at negative applied potentials, where the complete Z-scheme operates inefficiently. La co-doping fixes Rh in the 3+ state, which results in long-lived photogenerated electrons even at very positive potentials (+1 V versus the reversible hydrogen electrode), in which both components of the complete device operate effectively. This understanding of the role of co-dopants provides a new insight into the design principles for water-splitting devices based on bandgap-engineered metal oxides