Structures of 1∶1 inclusion complexes of cholic acid with fluoroethanols; Conformational analysis of fluoroethanols in the host cavity

The crystal structures of the inclusion complexes of cholic acid (CA) with 2-fluoroethanol (MFEtOH), 2,2-difluoroethanol (DFEtOH) and 2,2,2-trifluoroethanol (TFEtOH) have been determined by X-ray crystallography, which demonstrates that the guest alcohols are accommodated inside the cavity provided by CA molecules in a similar manner to that for the ethanol molecule in the CA-ethanol inclusion complex. As distinct from the ethanol molecule, the methylene C atoms of the fluoroethanols are not disordered; instead, the substituted F atoms are statistically disordered. All the alcohols are hydrogen-bonded to the OH groups of CA. The X-ray study showed that 46% of MFEtOH adopt thetrans-trans conformer, which is different from the exclusively predominant conformer in the gas and liquid phases, i.e.,thegauche-trans conformer. The study also showed that the F atoms of DFEtOH are statistically disordered, suggesting the possibility that three conformers exist inside the cavity. Such disorder presumably occurs in order to fill the vacant space around the CH₂F and CHF₂ groups inside the cavity. By contrast, we could not observe any disorder of the F atoms of TFEtOH. Supplementary Data relating to this article are deposited with the British Library as Supplementary Publication No. SUP 82176 (31 pages).     

Structure of synthetic transmembrane lipid membranes at the solid/liquid interface studied by specular X-ray reflectivity

We fabricated a new class of supported membranes based on monolayers of artificial bola (transmembrane) lipids. The lipids used in this study are symmetric bola lipids with two phosphocholine head groups, which resemble natural archaea lipids. To prevent bending of the hydrocarbon chains, stiff triple bonds are inserted in the middle of the hydrocarbon cores. The formation of homogeneous "monolayers" of transmembrane lipids over macroscopic areas can be monitored with fluorescence microscopy. Structures of such supported monolayers in bulk water were characterized with specular X-ray reflectivity using high energy X-ray radiation, which guarantees a high transmission through bulk water. Here, the vertical structure of single monolayers could be resolved from reconstructed electron density profiles. To verify the structural model suggested by the specular reflectivity, we also performed small- and wide-angle X-ray scattering of transmembrane lipid suspensions. The wide-angle patterns reflect a distorted chain-chain correlation, while the small-angle scattering allowed us to model an electron density profile which is consistent with the profile calculated from specular reflectivity.     

Live‐Cell Imaging of Endogenous mRNAs with a Small Molecule

Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific non‐engineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene‐specific RNA aptamer, combined with a cell‐permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live‐cell imaging of the endogenous mRNA of β‐actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin‐2, cortactin, and cytoplasmic FMR1‐interacting protein 2. The RNA‐imaging technology and its further optimization might permit live‐cell imaging of any RNA molecules.     

Self-assembly control of a pyridine-containing diblock copolymer by perfluorinated counter anions during salt-induced micellization

The micelle formation of poly[(4-pyridinemethoxymethyl)styrene]-block-polystyrene (PPySt-b-PSt) was studied in the nonselective solvent using perfluoroalkyl carboxylic acids. PPySt-b-PSt showed no self-assembly into micelles in THF, because this solvent was nonselective for the copolymer. Dynamic light scattering demonstrated that the diblock copolymer formed the micelles in the solvent in the presence of perfluoroalkyl carboxylic acids in which the number of carbons in the perfluoroalkyl chains was over eight. ¹H NMR revealed that the micellization proceeded through the salt formation of the pyridinium perfluoroalkyl carboxylate and through the aggregation of the perfluoroalkyl chains in the counter anions. The hydrodynamic radius and the aggregation number of the micelles increased with an increase in the length of the perfluoroalkyl chain. The copolymer needed less carboxylic acid with longer perfluoroalkyl chain to form the micelles. The copolymer produced the micelles with lower aggregation number and higher critical micelle concentration at higher temperature, although the micellar size was almost independent of the temperature. The micelles were unstable with respect to the variation in the temperature, and were dissociated into the unimers with the increase in the temperature. The micelles, however, were reconstructed by decreasing the temperature. This dissociation–reconstruction of the micelles was controlled reversibly not only by the temperature but also by the concentration of the perfluoroalkyl carboxylic acid. An increase in the acid concentration suppressed the dissociation into the unimers, while promoting the reconstruction of the micelles.