28 research outputs found
Nanoscale Confinement and Fluorescence Effects of Bacterial Light Harvesting Complex LH2 in Mesoporous Silicas
Many key chemical and biochemical reactions, particularly in living cells, take place in confined space at the mesoscopic scale. Toward understanding of physicochemical nature of biomacromolecules confined in nanoscale space, in this work we have elucidated fluorescence effects of a light harvesting complex LH2 in nanoscale chemical environments. Mesoporous silicas (SBA-15 family) with different shapes and pore sizes were synthesized and used to create nanoscale biomimetic environments for molecular confinement of LH2. A combination of UV-vis absorption, wide-field fluorescence microscopy, and in situ ellipsometry supports that the LH2 complexes are located inside the silica nanopores. Systematic fluorescence effects were observed and depend on degree of space confinement. In particular, the temperature dependence of the steady-state fluorescence spectra was analyzed in detail using condensed matter band shape theories. Systematic electronic-vibrational coupling differences in the LH2 transitions between the free and confined states are found, most likely responsible for the fluorescence effects experimentally observed
Prediction of Extracellular Proteases of the Human Pathogen Helicobacter pylori Reveals Proteolytic Activity of the Hp1018/19 Protein HtrA
Exported proteases of Helicobacter pylori (H. pylori) are potentially involved in pathogen-associated disorders leading to gastric inflammation and neoplasia. By comprehensive sequence screening of the H. pylori proteome for predicted secreted proteases, we retrieved several candidate genes. We detected caseinolytic activities of several such proteases, which are released independently from the H. pylori type IV secretion system encoded by the cag pathogenicity island (cagPAI). Among these, we found the predicted serine protease HtrA (Hp1019), which was previously identified in the bacterial secretome of H. pylori. Importantly, we further found that the H. pylori genes hp1018 and hp1019 represent a single gene likely coding for an exported protein. Here, we directly verified proteolytic activity of HtrA in vitro and identified the HtrA protease in zymograms by mass spectrometry. Overexpressed and purified HtrA exhibited pronounced proteolytic activity, which is inactivated after mutation of Ser205 to alanine in the predicted active center of HtrA. These data demonstrate that H. pylori secretes HtrA as an active protease, which might represent a novel candidate target for therapeutic intervention strategies
Environmental Acidification Drives S. pyogenes Pilus Expression and Microcolony Formation on Epithelial Cells in a FCT-Dependent Manner
Group A Streptococcus (GAS, Streptococcus pyogenes) is a Gram-positive human pathogen responsible for a diverse variety of diseases, including pharyngitis, skin infections, invasive necrotizing fasciitis and autoimmune sequelae. We have recently shown that GAS cell adhesion and biofilm formation is associated with the presence of pili on the surface of these bacteria. GAS pilus proteins are encoded in the FCT (Fibronectin- Collagen-T antigen) genomic region, of which nine different variants have been identified so far. In the present study we undertook a global analysis of GAS isolates representing the majority of FCT-variants to investigate the effect of environmental growth conditions on their capacity to form multicellular communities. For FCT-types 2, 3, 5 and 6 and a subset of FCT-4 strains, we observed that acidification resulting from fermentative sugar metabolism leads to an increased ability of the bacteria to form biofilm on abiotic surfaces and microcolonies on epithelial cells. The higher biofilm forming capacity at low environmental pH was directly associated with an enhanced expression of the genes encoding the pilus components and of their transcription regulators. The data indicate that environmental pH affects the expression of most pilus types and thereby the formation of multicellular cell-adhering communities that assist the initial steps of GAS infection
The Open, the Closed, and the Empty: Time-Resolved Fluorescence Spectroscopy and Computational Analysis of RC-LH1 Complexes from Rhodopseudomonas palustris
Comparison of the fluorescence kinetics of detergent -solubilized and membrane-reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides
Picosecond time-resolved fluorescence spectroscopy has been used in order to compare the fluorescence kinetics of detergent-solubilized and membrane-reconstituted light-harvesting 2 (LH2) complexes from the purple bacteria <i>Rhodopseudomonas</i> (<i>Rps.</i>) <i>acidophila</i> and <i>Rhodobacter</i> (<i>Rb.</i>) <i>sphaeroides</i>. LH2 complexes were reconstituted in phospholipid model membranes at different lipid:protein-ratios and all samples were studied exciting with a wide range of excitation densities. While the detergent-solubilized LH2 complexes from <i>Rps. acidophila</i> showed monoexponential decay kinetics (tf = 980 ps) for excitation densities of up to 3ú1013 photons/(pulseúcm2), the membrane-reconstituted LH2 complexes showed multiexponential kinetics even at low excitation densities and high lipid:protein-ratios. The latter finding indicates an efficient clustering of LH2 complexes in the phospholipid membranes. Similar results were obtained for the LH2 complexes from <i>Rb. sphaeroides</i>
1D Chains of Diruthenium Tetracarbonyl Sawhorse Complexes
The syntheses of five 1D coordination polymers containing the sawhorse‐type unit [Ru2(µ‐OAc)2(CO)4] linked by various bridging N,N‐donor ligands are reported. Various π‐conjugated linkers, such as trans‐[1,2‐bis(N‐methyl)imidazol‐2‐yl]ethylene (trans‐bie), pyrazine (pyz), 4,4′‐bipyridine (4,4′‐bipy) and 1,2‐bis(4‐pyridyl)ethylene (bpe) as well as the aliphatic linker 1,4‐diazabicyclo[2.2.2]octane (DABCO), were applied in the syntheses. The formation of 1D coordination polymers was proven by crystal structure determinations of two of the polymeric materials. The geometries and electronic structures of all polymers were analysed further by CASSCF/CASPT2 and DFT calculations based on monomeric model compounds. Finally, first attempts regarding deposition of the 1D chain, the sawhorse fragment and/or the ligands on a highly ordered pyrolytic graphite surface, analysed by STM measurements, are also reported