A solid-phase synthetic strategy for labeled peptides: synthesis of a biotinylated derivative of the delta opioid receptor antagonist TIPP (Tyr-Tic-Phe-Phe-OH)

A general solid-phase synthetic strategy for labeled peptides was developed and used to prepare a biotinylated derivative of the delta opioid receptor antagonist TIPP (Tyr-Tic-Phe-Phe-OH). A monoprotected hydrophilic diamine linker was attached to an aldehyde-containing solid-phase resin by reductive amination, followed by introduction of biotin and peptide synthesis to yield Tyr-Tic-Phe-Phe-Asp-NH(CH(2)CH(2)O)(2)CH(2)CH(2)NH-biotin (2). The high delta receptor affinity and selectivity of 2 demonstrate the applicability of this design approach for labeled peptide derivatives.     

Investigating the molecular mechanisms of in-plane mechanochemistry on cantilever arrays

Free-standing cantilevers, which directly translate specific biochemical reactions into micromechanical motion, have recently attracted much attention as label-free biosensors and micro/nano robotic devices. To exploit this mechanochemical sensing technology, it is essential to develop a fundamental understanding of the origins of surface stress. Here we report a detailed study into the molecular basis of stress generation in aqueous environments focusing on the pH titration of model mercaptohexadecanoic acid self-assembled monolayers (SAMs), using in situ reference cantilevers coated with nonionizable hexadecanethiol SAMs. Semiautomated data analysis and a statistical model were developed to quantify cyclic deprotonation/protonation reactions on multiple arrays. In-plane force titrations were found to have the sensitivity to detect ionic hydrogen bond formation between protonated and nonprotonated carboxylic acid groups in the proximity of the surface pK1/2, which generated a mean tensile differential surface stress of +1.2 +/- 0.3 mN/m at pH 6.0, corresponding to 1 pN attractive force between two adjacent MHA molecules. Conversely, the magnitude of compressive differential surface stress was found to increase progressively with pH >/= 7.0, reaching a maximum of -14.5 +/- 0.5 mN/m at pH 9.0, attributed to enhanced electrostatic repulsion between deprotonated carboxylic acid groups. However, striking differences were observed in the micromechanical responses to different ionic strength and ion species present in the aqueous environment, highlighting the critical role of counter- and co-ions on surface stress. Our findings provide fundamental insights into the molecular mechanisms of in-plane mechanochemistry, which may be exploited for biosensing and nanoactuation applications.     

Complexation of well-controlled low-molecular weight polyelectrolytes with antisense oligonucleotides

The influences of polymer-related properties such as molecular weight, charge density, counter ion, and hydrophilic block on the complexation of polyelectrolytes and a fluorescein-labeled oligonucleotide (ON) were investigated. A series of well-defined and well-controlled 2-(N,N-dimethylamino)ethyl methacrylate (DMAEMA) polymers and block copolymers were prepared using living anionic and radical polymerization methods. Fluorescence measurement was used to reveal the effects of polymer molecular weight, charge density, and counter ion type on the complexation. PolyDMAEMA samples having double molecular weights of the chosen oligonucleotide gave the optimal complexation performance. Kinetic studies showed that high-molecular weight/high-charge density polymer samples produced very stable complexes. The fully charged polyDMAEMA displayed the strongest binding with the ON. These complexes were therefore less sensitive to the changes in the environment. PolyDMAEMA–DMSQ samples had slightly higher complexation ability than polyDMAEMA–MCQ (DMSQ: dimethylsulfate quat; MCQ: methylchloride quat). Both poly(DMAEMA-b-HEMA) and poly(DMAEMA–MCQ-b-PEG) block copolymers showed good complexation ability and steric stability [HEMA: 2-hydroxyethyl methacrylate; PEG: poly(ethylene glycol)]. PEG, but not HEMA block, enhanced the effectiveness of polyDMAEMA–MCQ binding with the ON.     

Squeezing the [Cu-OH...H2O-Cu]3+ bridge by cryptate encapsulation

Treatment of cryptand L(1) with Cu(II) generates a H3O2(-)-bridged dicopper(II) cryptate, 2, where the guest anion has responded to steric constraint by a significant shortening of the O-O distance to 2.325(9) A; computational optimization at the B3LYP/6-31(d) level suggests that the bridging O-H...O H-bond is bent (approximately 157 degrees) but that the barrier to interchange of the bridging H atom is low (<4 kJ mol(-1)). This cryptate, rather than the [Cu2L(1)muCN]3+ species recently claimed to derive from cleavage of the C-C bond of the solvent, is the product of acetonitrile recrystallization of the initially formed reaction product, 1.     

Characterisation and proposed origin of mass spectrometric ions observed 30 Th above the ionised molecules of per-O-methylated carbohydrates

Derivatisation of carbohydrates by permethylation significantly improves the mass spectrometric intensity of carbohydrate-derived ions and allows more readily interpretable fragmentation; in addition, samples are conveniently separated from salts, and larger oligosaccharides are more readily ionised. It has previously been recognised that, in the mass spectra of permethylated carbohydrates, a series of ions indicating species 30 Da larger than the fully methylated carbohydrate molecules are also observed. These species have not been characterised in the literature despite their apparently ubiquitous occurrence in the mass spectra of permethylated carbohydrates. Tandem mass spectrometry (MS/MS) experiments were performed on permethylated carbohydrates and reduced permethylated carbohydrates that exhibit the artefact, demonstrating that the artefact is not reducing terminal specific, and that the artefact can be introduced at any hydroxyl residue. It was further demonstrated through the use of different alkylation reagents that the origin of this artefact group is the alkylating reagent itself. It is proposed that side reactions that occur between the permethylation reagents allow the production of small amounts of iodomethyl methyl ether. This reagent can then compete with methyl iodide for reaction with the carbohydrate -OH groups. The result is partial incorporation of a methoxymethyl moiety instead of a methyl group, detected as '+30' artefact ions.     

Temperature Dependence for Fluorescence of β-NADH in Glycerol/Water Solution and in Trehalose/Sucrose Glass

Fluorescence imaging of cells and tissue can be used to evaluate β-NADH redox and location. At low temperature, β-NADH fluorescence intensity increases and therefore sensitivity of imaging increases. In this paper, the temperature dependence of fluorescence was evaluated for β-NADH in glycerol/water solution and in trehalose/sucrose glass. The average fluorescence lifetime for NADH in glycerol/water is 0.66 ns, compared with 5.3 ns in trehalose/ sucrose at 20°C. Emission spectra were recorded from 290 to 12 K. The fluorescence of β-NADH in glycerol/water increases ∼16 fold and the emission shifts about 35 nm to the blue as temperature decreases. Much smaller change is seen for fluorescence of β-NADH in sugar glass. Below 77 K, the β-NADH spectral features did not change significantly with temperature change, and so no increase in sensitivity is obtained by going to very low temperatures. It is suggested that the sensitivity of β-NADH fluorescence is related to water relaxation around the excited state molecule. Differences in water in various tissues may contribute to β-NADH fluorescence changes when cells are altered.     

Effect of forewing and hindwing interactions on aerodynamic forces and power in hovering dragonfly flight

Dragonflies are four-winged insects that have the ability to control aerodynamic performance by modulating the phase lag (phi) between forewings and hindwings. We film the wing motion of a tethered dragonfly and compute the aerodynamic force and power as a function of the phase. We find that the out-of-phase motion as seen in steady hovering uses nearly minimal power to generate the required force to balance the weight, and the in-phase motion seen in takeoffs provides an additional force to accelerate. We explain the main hydrodynamic interaction that causes this phase dependence.     

Surface characterization of synthetic and mineral carbonate fluoroapatites

As part of a larger project concerning the sorption capacities of carbonate fluoroapatites, the physical and surface properties of the synthetic carbonate fluoroapatite and mineral francolite retained for this work have been carefully examined. The average particle size of the powdered samples, determined by laser granulometry, is around 30 microm. Their specific surface areas, measured according to the N(2) BET method, are 8.8+/-0.1 and 13.9+/-0.1 m(2).g(-1), respectively, for the synthetic and the mineral compound. The solubility of both solids has been studied under undersaturation initial conditions. The synthetic compound has a solubility product of 10(-103+/-2), but the mineral francolite, which is highly substituted and poorly crystalline, is more soluble and dissolves incongruently. The amphoteric properties of the hydrous carbonate fluoroapatites surfaces have also been investigated through zeta-potential measurements and potentiometric titrations in KNO(3) medium at 25 degrees C. The experimental data have been interpreted using the computer program FITEQL 3.2, applying either the nonelectrostatic or the constant capacitance model. For both solids, the values of the surface acidity constants determined with the two models are very close, suggesting a weak contribution of the electrostatic interactions. Moreover, the more acidic surface of the synthetic compound compared to that of the mineral one is assigned to the differences in crystallinity and chemical composition between the two solids.     

Properties of the liquid crystals formed by certain 4'-n-alkylbiphenyl-4-yl 5-n-alkylthiophene-2-carboxylates

Thirty compounds, ten members of each of three homologous series of esters derived from 4-hydroxy-4'-n-octyl-, -nonyl-, and -decyl-biphenyl and the 5-n-alkylthiophene-2-carboxylic acids (methyl through to decyl homologues) have been prepared. The liquid crystal properties of these esters, which show extensive smectic polymorphism, have been investigated by thermal optical microscopy, differential scanning calorimetry, and miscibility studies. Four members of the corresponding 4-n-alkylbenzoates have also been prepared in order to compare their liquid crystal behaviour with the 5-n-alkylthiophene-2-carboxylate counterparts: both smectic and nematic thermal stability are higher for the 4-n-alkylbenzoates, by 30.6° and 36.8° C, respectively.