Johannes Kepler University of Linz

About: Johannes Kepler University of Linz is a education organization based out in Linz, Oberösterreich, Austria. It is known for research contribution in the topics: Thin film & Quantum dot. The organization has 6605 authors who have published 19243 publications receiving 385667 citations.

Structure, cell wall elasticity and polysaccharide properties of living yeast cells, as probed by AFM

Abstract: Although the chemical composition of yeast cell walls is known, the organization, assembly, and interactions of the various macromolecules remain poorly understood. Here, we used in situ atomic force microscopy (AFM) in three different modes to probe the ultrastructure, cell wall elasticity and polymer properties of two brewing yeast strains, i.e. Saccharomyces carlsbergensis and S. cerevisiae. Topographic images of the two strains revealed smooth and homogeneous cell surfaces, and the presence of circular bud scars on dividing cells. Nanomechanical measurements demonstrated that the cell wall elasticity of S. carlsbergensis is homogeneous. By contrast, the bud scar of S. cerevisiae was found to be stiffer than the cell wall, presumably due to the accumulation of chitin. Notably, single molecule force spectroscopy with lectin-modified tips revealed major differences in polysaccharide properties of the two strains. Polysaccharides were clearly more extended on S. cerevisiae, suggesting that not only oligosaccharides, but also polypeptide chains of the mannoproteins were stretched. Consistent with earlier cell surface analyses, these findings may explain the very different aggregation properties of the two organisms. This study demonstrates the power of using multiple complementary AFM modalities for probing the organization and interactions of the various macromolecules of microbial cell walls.

Photosystem II does not possess a simple excitation energy funnel: time-resolved fluorescence spectroscopy meets theory.

Abstract: The experimentally obtained time-resolved fluorescence spectra of photosystem II (PS II) core complexes, purified from a thermophilic cyanobacterium Thermosynechococcus vulcanus, at 5-180 K are compared with simulations. Dynamic localization effects of excitons are treated implicitly by introducing exciton domains of strongly coupled pigments. Exciton relaxations within a domain and exciton transfers between domains are treated on the basis of Redfield theory and generalized Forster theory, respectively. The excitonic couplings between the pigments are calculated by a quantum chemical/electrostatic method (Poisson-TrEsp). Starting with previously published values, a refined set of site energies of the pigments is obtained through optimization cycles of the fits of stationary optical spectra of PS II. Satisfactorily agreement between the experimental and simulated spectra is obtained for the absorption spectrum including its temperature dependence and the linear dichroism spectrum of PS II core complexes (PS II-CC). Furthermore, the refined site energies well reproduce the temperature dependence of the time-resolved fluorescence spectrum of PS II-CC, which is characterized by the emergence of a 695 nm fluorescence peak upon cooling down to 77 K and the decrease of its relative intensity upon further cooling below 77 K. The blue shift of the fluorescence band upon cooling below 77 K is explained by the existence of two red-shifted chlorophyll pools emitting at around 685 and 695 nm. The former pool is assigned to Chl45 or Chl43 in CP43 (Chl numbering according to the nomenclature of Loll et al. Nature2005, 438, 1040) while the latter is assigned to Chl29 in CP47. The 695 nm emitting chlorophyll is suggested to attract excitations from the peripheral light-harvesting complexes and might also be involved in photoprotection.

Diffusion-Separated Nuclear Magnetic Resonance Spectroscopy of Polymer Mixtures

Abstract: The potential of diffusion-ordered 2D NMR spectroscopy (DOSY) for the analysis of solutions of polymer mixtures and polymers with complex molecular mass distributions is investigated. Diffusion coefficient labeling in NMR is generally achieved by stepwise ramping up of the amplitudes of pairs of pulsed field gradients (PFGs). After Fourier transformation in the acquisition dimension and an inverse Laplace transform (ILT) with respect to the square of the gradient strength, 2D spectra are obtained that show the chemical shift along one dimension and the translational diffusion coefficient along the other. Since polymers may have broad, nonsymmetric or complex (bi- and multimodal) molecular weight distributions (MWDs), the diffusion coefficient distribution should follow the main features of the MWD. However, the calculation of the diffusion coefficient distribution involves a numerically unstable data inversion (ILT), which limits the resolution in the diffusion dimension. The applications of DOSY NMR tech...