Structural Coupling of 11‐cis‐7‐Methyl‐retinal and Amino Acids at the Ligand Binding Pocket of Rhodopsin†

It was previously shown that opsin can be regenerated with the newly synthesized 11‐cis‐7‐methyl‐retinal forming an artificial visual pigment. We now extend this study to include mutants at positions close to the retinal to further dissect the interactions of native and artificial chromophores with opsin. Several mutants at M207, W265 and Y268 have been obtained and regenerated with 11‐cis‐retinal and the 7‐methyl analog. M207 is the site of the point mutation M207R associated with the retinal degenerative disease retinitis pigmentosa. All the studied mutants regenerated with 11‐cis‐retinal except for M207C which proved to be completely misfolded. The naturally occurring M207R mutant formed a pigment with an unprotonated Schiff base linkage, altered photobleaching and low MetarhodopsinII stability. Mutants regenerated with the 7‐methyl analog showed altered photobleaching reflecting a structural perturbation in the vicinity of M207. The newly obtained mutants at M207 also showed reduced levels of transducin activation with M207R showing essentially no transducin activation. Our results highlight the tight coupling of the vicinity of C7 of retinal and M207 and support the involvement of this amino acid residue in the conformational changes associated with rhodopsin photoactivation.     

Structural Behavior of Pt–Sn Supported on MgO

Pt–Sn supported on magnesia and alumina were characterized, before and after treatment with hydrogen, by Mössbauer spectroscopy and X-ray diffraction. For the calcined samples on both supports tin is present as SnO₂ and platinum as metal. After reduction with hydrogen, platinum and tin diffuse into the magnesia lattice to form a solid solution. On alumina Sn(IV), Sn(II), Sn(0), Pt, Pt₃Sn, PtSn and PtSn₂ alloys are formed. The SnO interacts strongly with the alumina support. The catalytic activity of both Pt–Sn catalysts is strongly affected by the support. On alumina the dehydrogenation of cyclohexane is very high, whereas that on magnesia is almost non-active.     

Structural Characterization of a Zinc High‐affinity Binding Site in Rhodopsin†

For the first time to our knowledge, X‐ray absorption spectroscopy (XAS) has been used to investigate the environment of putative Zn²⁺ binding sites in rhodopsin. We studied native purified nondeionized rhodopsin without any further addition of Zn²⁺, as well as with 1.5 mol of Zn²⁺—as zinc chloride—per mole of protein. Three different binding sites in rhodopsin were considered based on computational chemistry studies, and a quantitative analysis of the XAS signal was performed by fitting the experimental data to their simulated XAS spectra. Our results demonstrate that Zn²⁺ is intrinsically bound to rhodopsin and are compatible with the existence of an octahedral coordination involving six oxygen atoms in the first shell (average Zn‐O distance of 2.08 Å), and with a second coordination shell containing one or two phosphorus or sulfur atoms at an average distance of 2.81 Å.     

Comparative analysis of glycosylinositol phosphorylceramides from fungi by electrospray tandem mass spectrometry with low-energy collision-induced dissociation of Li(+) adduct ions.

Glycosylinositol phosphorylceramides (GIPCs) are a class of acidic glycosphingolipids (GSLs) expressed by fungi, plants, and certain parasitic organisms, but not found in cells or tissues of mammals or other higher animals. Recent characterizations of fungal GIPCs point to an emerging diversity which could rival that already known for mammalian GSLs, and which can be expected to present a multitude of challenges for the analytical chemist. Previously, the use of Li(+) cationization, in conjunction with electrospray ionization mass spectrometry (ESI-MS) and low-energy collision-induced dissociation tandem mass spectrometry (ESI-MS/CID-MS), was found to be particularly effective for detailed structural analysis of monohexosylceramides (cerebrosides) from a variety of sources, including fungi, especially minor components present in mixtures at extremely low abundance. In applying Li(+) cationization to characterization of GIPCs, a substantial increase in both sensitivity and fragmentation was observed on collision-induced dissociation of [M + Li](+) versus [M + Na](+) for the same components analyzed under similar conditions, similar to results obtained previously with cerebrosides. Molecular adduct fragmentation patterns were found to be systematic and characteristic for both the glycosylinositol and ceramide moieties with or without phosphate. Interestingly, significant differences were observed in fragmentation patterns when comparing GIPCs having Manalpha1 --> 2 versus Manalpha1 --> 6Ins core linkages. In addition, it was useful to perform tandem product ion scans on primary fragments generated in the orifice region, equivalent to ESI-(CID-MS)(2) mode. Finally, precursor ion scanning from appropriate glycosylinositol phosphate product ions yielded clean molecular ion profiles in the presence of obscuring impurity peaks. The methods were applied to detailed characterization of GIPC fractions of increasing structural complexity from a variety of fungi, including a non-pathogenic Basidiomycete (mushroom), Agaricus blazei, and pathogenic Euascomycete species such as Aspergillus fumigatus, Histoplasma capsulatum, and Sporothrix schenckii. The analysis confirmed a remarkable diversity of GIPC structures synthesized by the dimorphic S. schenckii, as well as differential expression of both glycosylinositol and ceramide structures in the mycelium and yeast forms of this mycopathogen. Mass spectrometry also established that the ceramides of some A. fumigatus GIPC fractions contain very little 2-hydroxylation of the long-chain fatty-N-acyl moiety, a feature that is not generally observed with fungal GIPCs.     

Porous,bicontinuous, and cationic polyelectrolyte obtained by high internal phase emulsion polymerization

In this work, a cationic polyelectrolyte was synthesized through oil‐in‐water high internal phase emulsion polymerization. The porous polymer was obtained using the monomer (4‐vinyl benzyl)trimethylammonium chloride and cross‐linked with N,N‐methylene‐bis‐acrylamide; additionally, ethylene glycol dimethacrylate (EGDMA) was used as the second cross‐linker, which was solubilized in the discontinuous phase leading to a bicontinuous‐like HIPE system because of the characteristics of this cross‐linker and the phase, where polymerized several effects on the polyHIPE were expected. In this way, the effect of the emulsifier and EGDMA content on the pore size, swelling, and rheological properties was assessed. It was observed that an increased concentration of the emulsifier in the polymerization decreased the pore size, narrowed its size distribution, and diminished the swelling capacity of the polymer. Additionally, the poly (HIPE) displayed a close‐cell structure explained by the locus of initiation starting from the droplets of the emulsion. After the addition of EGDMA, the polymer exhibited a major decrease in pore size and a significant decrease in swelling attributed to the polymerized skin layer on the droplet and hydrophobicity provided by the polyEGDMA, respectively. Rheological assays revealed an increase in the complex modulus and shear stress as the pore size decreased, but the addition of EGDMA did not produce an increase in the modulus, as expected. Finally, the sorption capabilities of the cationic porous polymers were evaluated through kinetic and isotherm sorption experiments using the anionic dye Acid Black 24.     

Histochemical and ultrastructural study of collagen fibers in mouse pubic symphysis during late pregnancy

Reference is usually made to the parallel orientation towards the main line of exerted tension at the pubic joint in mice, for supporting forces applied to the joint. Despite the wealth of morphological information about the extracellular matrix in this joint, little is known regarding the involvement of the crimp of collagen fibers in the dramatic transformations occurring in this region during the last 3 days of pregnancy. Examination of the collagenous architecture suggests that the biomechanical properties are directly related to fibril diameters, composition of ground substance and changes in the bundle morphology, particularly in the crimp structure. The purpose of this study was to further describe the transformation of the collagen fibers of the pubic symphysis during late mouse pregnancy. We examined the architecture of collagen fibers in the symphysis and pubic ligament through the Picrosirius-polarization method and also through scanning electron microscopy to directly visualize and measure the crimping from pregnant and virgin mice. The crimp angle and the length of five consecutive crimps were measured according to Patterson-Kane et al. [Connect. Tissue Res. 36 (1997) 253]. It could be demonstrated that the angles progressively decreased and the crimp length increased, denoting that the fibers have untwisted during the relaxation process. Our findings suggest that a disruption of the helical arrangement of the collagen containing fibers may contribute to explaining the rapid remodeling that occurs at the end of pregnancy and that is responsible for an increase in pliancy and length of the pubic ligament in mice.     

Novel cross-coupling reactions between organotellurides and Grignard reagents employing a MnCl2/CuI catalytic system

We present a general protocol for the cross-coupling reaction of Grignard reagents and organic tellurides. Aryl Grignard reagents react stereospecifically with vinyl tellurides in the presence of a catalytic amount of manganese (II) chloride and copper (I) iodide to produce good yields of the corresponding cross-coupling products.     

A Grothendieck-Riemann-Roch formula for maps of complex manifolds

Mathematische Annalen -     

Collisional spreading of small amplitude nuclear vibrations

We present a microscopic treatment of the spreading widths of giant resonances — a collisional RPA (with complex frequencies,z=iω+γ) which turns out to have a very simple analytical form. Its essential ingredients are:a) The usual mean field contribution, which reproduces the frequencies of the collisionless RPA andb) non-unitary (collision) contributions which arise from the inclusion of quantum correlation effects in the weak coupling approximation. This collisional RPA equation can be easily put to quantitative use in realistic nuclear structure problems.