Novel Xylene‐Linked Maltoside Amphiphiles (XMAs) for Membrane Protein Stabilisation

Membrane proteins are key functional players in biological systems. These biomacromolecules contain both hydrophilic and hydrophobic regions and thus amphipathic molecules are necessary to extract membrane proteins from their native lipid environments and stabilise them in aqueous solutions. Conventional detergents are commonly used for membrane protein manipulation, but membrane proteins surrounded by these agents often undergo denaturation and aggregation. In this study, a novel class of maltoside‐bearing amphiphiles, with a xylene linker in the central region, designated xylene‐linked maltoside amphiphiles (XMAs) was developed. When these novel agents were evaluated with a number of membrane proteins, it was found that XMA‐4 and XMA‐5 have particularly favourable efficacy with respect to membrane protein stabilisation, indicating that these agents hold significant potential for membrane protein structural study.     

The dehydration of SrTeO3(H2O)--a topotactic reaction for preparation of the new metastable strontium oxotellurate(IV) phase ε-SrTeO3

Microcrystalline single-phase strontium oxotellurate(IV) monohydrate, SrTeO(3)(H(2)O), was obtained by microwave-assisted hydrothermal synthesis under alkaline conditions at 180 °C for 30 min. A temperature of 220 °C and longer reaction times led to single crystal growth of this material. The crystal structure of SrTeO(3)(H(2)O) was determined from single crystal X-ray diffraction data: P2(1)/c, Z = 4, a = 7.7669(5), b = 7.1739(4), c = 8.3311(5) ?, β = 107.210(1)°, V = 443.42(5) ?(3), 1403 structure factors, 63 parameters, R[F(2)>2σ(F(2))] = 0.0208, wR(F(2) all) = 0.0516, S = 1.031. SrTeO(3)(H(2)O) is isotypic with the homologous BaTeO(3)(H(2)O) and is characterised by a layered assembly parallel to (100) of edge-sharing [SrO(6)(H(2)O)] polyhedra capped on each side of the layer by trigonal-prismatic [TeO(3)] units. The cohesion of the structure is accomplished by moderate O-H···O hydrogen bonding interactions between donor water molecules and acceptor O atoms of adjacent layers. In a topochemical reaction, SrTeO(3)(H(2)O) condensates above 150 °C to the metastable phase ε-SrTeO(3) and transforms upon further heating to δ-SrTeO(3). The crystal structure of ε-SrTeO(3), the fifth known polymorph of this composition, was determined from combined electron microscopy and laboratory X-ray powder diffraction studies: P2(1)/c, Z = 4, a = 6.7759(1), b = 7.2188(1), c = 8.6773(2) ?, β = 126.4980(7)°, V = 341.20(18) ?(3), R(Fobs) = 0.0166, R(Bobs) = 0.0318, Rwp = 0.0733, Goof = 1.38. The structure of ε-SrTeO(3) shows the same basic set-up as SrTeO(3)(H(2)O), but the layered arrangement of the hydrous phase transforms into a framework structure after elimination of water. The structural studies of SrTeO(3)(H(2)O) and ε-SrTeO(3) are complemented by thermal analysis and vibrational spectroscopic measurements.     

Immobilization of molecularly imprinted polymer nanoparticles in electrospun poly(vinyl alcohol) nanofibers

We describe the fabrication of polymer nanofibers with entrapped molecularly imprinted polymer (MIP) nanoparticles and study their possible use in a fluorescence-based biosensor application. The MIP was imprinted with the fluorescent amino acid derivative dansyl-L-phenylalanine. Poly(vinyl alcohol) was used as a support for MIP nanoparticles because it is water-soluble and can be spun into very thin fibers. The fibers were characterized by atomic force microscopy and optical microscopy, and fluorescence microscopy was used for the characterization of target binding to the MIP. The fibers show close to 100% recovery upon extraction and rebinding of the target molecule. The selectivity of the system has been demonstrated through competitive binding experiments with nonfluorescent analogues boc-L-phenylalanine and boc-D-phenylalanine.     

Bifunctional co-initiators: A new strategy for the design of efficient systems in radical photopolymerization reactions under air

Bifunctional co-initiators containing amine and silane groups (silylamines) are proposed for radical photopolymerization in experimental conditions (e.g. low light intensity, low viscosity thin samples, UV and visible light exposure) where a strong deleterious oxygen effect occurs. In the presence of benzophenone, isopropylthioxanthone, camphorquinone, 2,6-diphenyl-4-(4′-dimethylaminophenyl) thiopyrylium tetrafluoroborate, chloro-HABI and cleavable photoinitiators, these compounds show a high reactivity under air, even better than that observed for a reference amine as a hydrogen donor. The associated mechanisms are investigated by laser flash photolysis (LFP) and Kinetic ESR.     

Accessible Mannitol‐Based Amphiphiles (MNAs) for Membrane Protein Solubilisation and Stabilisation

Integral membrane proteins are amphipathic molecules crucial for all cellular life. The structural study of these macromolecules starts with protein extraction from the native membranes, followed by purification and crystallisation. Detergents are essential tools for these processes, but detergent‐solubilised membrane proteins often denature and aggregate, resulting in loss of both structure and function. In this study, a novel class of agents, designated mannitol‐based amphiphiles (MNAs), were prepared and characterised for their ability to solubilise and stabilise membrane proteins. Some of MNAs conferred enhanced stability to four membrane proteins including a G protein‐coupled receptor (GPCR), the β₂ adrenergic receptor (β₂AR), compared to both n‐dodecyl‐d ‐maltoside (DDM) and the other MNAs. These agents were also better than DDM for electron microscopy analysis of the β₂AR. The ease of preparation together with the enhanced membrane protein stabilisation efficacy demonstrates the value of these agents for future membrane protein research.     

Mesitylene‐Cored Glucoside Amphiphiles (MGAs) for Membrane Protein Studies: Importance of Alkyl Chain Density in Detergent Efficacy

Detergents serve as useful tools for membrane protein structural and functional studies. Their amphipathic nature allows detergents to associate with the hydrophobic regions of membrane proteins whilst maintaining the proteins in aqueous solution. However, widely used conventional detergents are limited in their ability to maintain the structural integrity of membrane proteins and thus there are major efforts underway to develop novel agents with improved properties. We prepared mesitylene‐cored glucoside amphiphiles (MGAs) with three alkyl chains and compared these agents with previously developed xylene‐linked maltoside agents (XMAs) with two alkyl chains and a conventional detergent (DDM). When these agents were evaluated for four membrane proteins including a G protein‐coupled receptor (GPCR), some agents such as MGA‐C13 and MGA‐C14 resulted in markedly enhanced stability of membrane proteins compared to both DDM and the XMAs. This favourable behaviour is due likely to the increased hydrophobic density provided by the extra alkyl chain. Thus, this study not only describes new glucoside agents with potential for membrane protein research, but also introduces a new detergent design principle for future development.