Computational design of transmembrane pores.

Transmembrane channels and pores have key roles in fundamental biological processes1 and in biotechnological applications such as DNA nanopore sequencing2-4, resulting in considerable interest in the design of pore-containing proteins. Synthetic amphiphilic peptides have been found to form ion channels5,6, and there have been recent advances in de novo membrane protein design7,8 and in redesigning naturally occurring channel-containing proteins9,10. However, the de novo design of stable, well-defined transmembrane protein pores that are capable of conducting ions selectively or are large enough to enable the passage of small-molecule fluorophores remains an outstanding challenge11,12. Here we report the computational design of protein pores formed by two concentric rings of α-helices that are stable and monodisperse in both their water-soluble and their transmembrane forms. Crystal structures of the water-soluble forms of a 12-helical pore and a 16-helical pore closely match the computational design models. Patch-clamp electrophysiology experiments show that, when expressed in insect cells, the transmembrane form of the 12-helix pore enables the passage of ions across the membrane with high selectivity for potassium over sodium; ion passage is blocked by specific chemical modification at the pore entrance. When incorporated into liposomes using in vitro protein synthesis, the transmembrane form of the 16-helix pore-but not the 12-helix pore-enables the passage of biotinylated Alexa Fluor 488. A cryo-electron microscopy structure of the 16-helix transmembrane pore closely matches the design model. The ability to produce structurally and functionally well-defined transmembrane pores opens the door to the creation of designer channels and pores for a wide variety of applications

Fabrication of Au/Pd alloy nanoparticle/Pichia pastoris composites: a microorganism-mediated approach

Fundamental Research Funds for Central Universities [2010121051]; NSFC projects [21106117, 21036004]Synthesis of metal nanoparticles (NPs) is in the limelight in modern nanotechnology. In this present study, bimetallic Au/Pd NP/Pichia pastoris composites were successfully fabricated through a one-pot microbial reduction of aqueous HAuCl4 and PdCl2 in the presence of H-2 as an electron donor. Interestingly, flower-like alloy Au/Pd NP/Pichia pastoris composites were obtained under the following conditions, NaCl concentration 0.9% (w/v), molar ratio of Au/Pd (1 : 2) and the time for pre-adsorption of Au(III) and Pd(II) ions 15 min, through fresh yeast reduction. The mapping results from scanning transmission electron microscopy (STEM) with a high-angle annular dark field detector confirmed that the Au/Pd NPs on the surface of the yeast were indeed alloy. Furthermore, the energy dispersive X-ray (EDX) and X-ray photoelectron spectroscopy (XPS) measurements showed that the composition of the bimetallic NPs were consistent with the initial molar ratio of the precursors

Biogenic flower-shaped Au-Pd nanoparticles: Synthesis, SERS detection and catalysis towards benzyl alcohol oxidation

～40 nm flower-shaped Au-Pd bimetallic nanoparticles were prepared in a facile and eco-friendly way based on the simultaneous bioreduction of HAuCl 4 and Na2PdCl4 with ascorbic acid and Cacumen Platycladi leaf extract at room temperature. Characterization techniques, such as transmission electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction, were employed to confirm that the as-synthesized nanoparticles were alloys. The obtained flower-shaped Au-Pd alloy nanoparticles exhibited an excellent surface enhanced Raman spectroscopic activity with rhodamine 6G and efficient catalytic ability for the oxidation of benzyl alcohol to benzaldehyde. ? 2014 The Royal Society of Chemistry

Pain-causing stinging nettle toxins target TMEM233 to modulate NaV1.7 function

Voltage-gated sodium (NaV) channels are critical regulators of neuronal excitability and are targeted by many toxins that directly interact with the pore-forming α subunit, typically via extracellular loops of the voltage-sensing domains, or residues forming part of the pore domain. Excelsatoxin A (ExTxA), a pain-causing knottin peptide from the Australian stinging tree Dendrocnide excelsa, is the first reported plant-derived NaV channel modulating peptide toxin. Here we show that TMEM233, a member of the dispanin family of transmembrane proteins expressed in sensory neurons, is essential for pharmacological activity of ExTxA at NaV channels, and that co-expression of TMEM233 modulates the gating properties of NaV1.7. These findings identify TMEM233 as a previously unknown NaV1.7-interacting protein, position TMEM233 and the dispanins as accessory proteins that are indispensable for toxin-mediated effects on NaV channel gating, and provide important insights into the function of NaV channels in sensory neurons

Interaction between graphene-coated nanowires revisited with transformation optics

The interaction between graphene-coated nanostructures provides interesting optical properties not found in isolated graphene plasmonic structures. However, full-analytical solutions, which can provide deep physical insights underlying the hybrid graphene plasmonic systems, are difficult to achieve. In this Letter, we deploy the theory of transformation optics to study the plasmonic interactions between two dielectric-core-graphene-shell nanowires. The scattering and absorption spectra as well as the field distributions are derived analytically. We find that the interaction between two graphene-coated nanowires results in polarization-independent multi-frequency Fano dips, which show a broadband red shift of bonding modes and a blue shift of anti-bonding modes when the nanowires approach each other. The analytical tool presented here offers a rigorous study of graphene plasmonic compound and can be extended to treat more complicated cases.NRF (Natl Research Foundation, S’pore)MOE (Min. of Education, S’pore)Accepted versio

Marking the status and development of marine VOCs recovery technology

During the storage and transportation of crude oil, volatile organic vapors (VOCs) are generated. International organizations and relevant countries have formulated relevant regulations for VOCs emissions. This article introduces four kinds of oil and gas recovery technologies and their combined processes: absorption method, adsorption method, membrane separation method and condensation method. The advantages and disadvantages of different oil and gas recovery technologies are analyzed and the process performance is compared. It lists examples of the application of oil and gas recovery technology in ships, points out its shortcomings, and puts forward relevant suggestions for the research of marine VOCs recovery technology

Raptor couples mTORC1 and ERK1/2 inhibition by cardamonin with oxidative stress induction in ovarian cancer cells

Background A balance on nutrient supply and redox homeostasis is required for cell survival, and increased antioxidant capacity of cancer cells may lead to chemotherapy failure. Objective To investigate the mechanism of anti-proliferation of cardamonin by inducing oxidative stress in ovarian cancer cells. Methods After 24 h of drug treatment, CCK8 kit and wound healing test were used to detect cell viability and migration ability, respectively, and the ROS levels were detected by flow cytometry. The differential protein expression after cardamonin administration was analyzed by proteomics, and the protein level was detected by Western blotting. Results Cardamonin inhibited the cell growth, which was related to ROS accumulation. Proteomic analysis suggested that MAPK pathway might be involved in cardamonin-induced oxidative stress. Western blotting showed that cardamonin decreased Raptor expression and the activity of mTORC1 and ERK1/2. Same results were observed in Raptor KO cells. Notably, in Raptor KO cells, the effect of cardamonin was weakened. Conclusion Raptor mediated the function of cardamonin on cellular redox homeostasis and cell proliferation through mTORC1 and ERK1/2 pathways

The Key Roles of GSK-3β in Regulating Mitochondrial Activity

Glycogen synthase kinase-3β (GSK-3β), a serine/threonine protein kinase, has been reported to show essential roles in molecular pathophysiology of many diseases. Mitochondrion is a dynamic organelle for producing cellular energy and determining cell fates. Stress-induced translocated GSK-3β may interact with mitochondrial proteins, including PI3K-Akt, PGC-1α, HK II, PKCε, components of respiratory chain, and subunits of mPTP. Mitochondrial pool of GSK-3β has been implicated in mediation of mitochondrial functions. GSK-3β exhibits the regulatory effects on mitochondrial biogenesis, mitochondrial bioenergetics, mitochondrial permeability, mitochondrial motility, and mitochondrial apoptosis. The versatile functions of GSK-3β might be associated with its wide range of substrates. Accumulative evidence demonstrates that GSK-3β inactivation may be potentially developed as the promising strategy in management of many diseases, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). Intensive efforts have been made for exploring GSK-3β inhibitors. Natural products provide us a great source for screening new lead compounds in inactivation of GSK-3β. The key roles of GSK-3β in mediation of mitochondrial functions are discussed in this review

Rape Pollen-Templated Synthesis of C,N Self-Doped Hierarchical TiO₂ for Selective Hydrogenation of 1,3-Butadiene

Herein, we described a mild and facile method for the synthesis of three-dimensional (3D) hierarchically porous TiO₂. This method simultaneously realized the C,N self-dopant, introduced more surface defectives, and decreased in the band-gap energy (<i>E</i>_g) of TiO₂. Rape pollen was chosen as the biotemplate to prepare TiO₂, and the pore property of the obtained products could be adjusted by changing the amount of Ti precursor and calcination temperature. The as-synthesized pollen-TiO₂ not only duplicated the morphology of rape pollen, but also showed enhanced 1-butene selectivity in the reaction of selective hydrogenation of 1,3-butadiene after loading elemental Pd, when comparing with the commercial TiO₂. Butenes selectivity of 98% and 1-butene selectivity of 66% were achieved at 50 °C, because of the hierarchically porous structure of pollen-TiO₂ and the lower <i>E</i>_g and more surface defects resulting from the C,N dopant