High resolution spectroscopy reveals fibrillation inhibition pathways of insulin

Fibril formation implies the conversion of a protein’s native secondary structure and is associated with several neurodegenerative diseases. A better understanding of fibrillation inhibition and fibril dissection requires nanoscale molecular characterization of amyloid structures involved. Tip-enhanced Raman scattering (TERS) has already been used to chemically analyze amyloid fibrils on a sub-protein unit basis. Here, TERS in combination with atomic force microscopy (AFM), and conventional Raman spectroscopy characterizes insulin assemblies generated during inhibition and dissection experiments in the presence of benzonitrile, dimethylsulfoxide, quercetin, and β-carotene. The AFM topography indicates formation of filamentous or bead-like insulin self-assemblies. Information on the secondary structure of bulk samples and of single aggregates is obtained from standard Raman and TERS measurements. In particular the high spatial resolution of TERS reveals the surface conformations associated with the specific agents. The insulin aggregates formed under different inhibition and dissection conditions can show a similar morphology but differ in their β-sheet structure content. This suggests different aggregation pathways where the prevention of the β-sheet stacking of the peptide chains plays a major role. The presented approach is not limited to amyloid-related reasearch but can be readily applied to systems requiring extremely surface-sensitive characterization without the need of labels

Label-free monitoring of plasmonic catalysis on the nanoscale

Plasmonics is the description of specific light matter interactions of metallic structures. In general the size of such structures is well in the nanometer regime and also determines such specific characteristics as color, field confinement etc. Plasmon-induced hot electrons play a vital role in so-called plasmonic catalysis, a field that has recently attracted attention as a new reaction platform. Current reports introduce such nanoscale catalysis as an effective approach to concentrate the energy of visible light and direct it to adsorbed molecules, thereby increasing the chemical reaction rate, and controlling the reaction selectivity. In this review, we present various plasmon-catalyzed reactions specifically monitored with Raman spectroscopy, namely surface-enhanced Raman scattering (SERS), remote SERS (Re-SERS) and tip-enhanced Raman scattering (TERS). These techniques utilize the signal enhancing effect of the metal nanoparticles. However, at the same time they can be used to control the actual reactivity. In the first part, the mechanism of plasmonic catalysis is introduced. Then it is shown how catalytic reactions can be spectroscopically investigated far beyond the diffraction limit using TERS. Finally, the sensitivity of the methods is discussed

Single molecule level plasmonic catalysis – a dilution study of p-nitrothiophenol on gold dimers

Surface plasmons on isolated gold dimers can initiate intermolecular reactions of adsorbed p-nitrothiophenol. At the single molecule level when dimerization is not possible an intramolecular reaction can be observed. Experimental evidence indicates that plasmon-induced hot electrons provide the required activation energy

Plasmon induced deprotonation of 2-mercaptopyridine

Surface plasmons can provide a novel route to induce and simultaneously monitor selective bond formation and breakage. Here pH-induced protonation, followed by plasmon-induced deprotonation of 2-mercaptopyridine was investigated using surface- and tip-enhanced Raman scattering (SERS and TERS). A large difference in the deprotonation rate between SERS and TERS will be demonstrated and discussed with respect to hot-spot distribution. © 2020 The Royal Society of Chemistry

Light-trapping structures for planar solar cells inspired by transformation optics

Optimal light absorption is decisive in obtaining high-efficiency solar cells. An established, if not to say the established, approach is to texture the interface of the light-absorbing layer with a suitable microstructure. However, structuring the light-absorbing layer is detrimental concerning its electrical properties due to an increased surface recombination rate (owing to enlarged surface area and surface defects) caused by the direct patterning process itself. This effect lowers the efficiency of the final solar cells. To circumvent this drawback, this work theoretically explores a transformation optics (TrO) inspired approach to map the nanopatterned texture onto a planar equivalent. This offers a pattern with the same optical functionality but with much improved electrical properties. Schwarz-Christoffel mappings are used for ensuring conformality of the maps. It leads to planar, inhomogeneous, dielectric-only materials for the light trapping structure to be placed on top of the planar light-absorbing layer. Such a design strategy paves a way towards a novel approach for implementing light-trapping structures into planar solar cells

Detection of Protein Glycosylation Using Tip-Enhanced Raman Scattering

The correct glycosylation of biopharmaceutical glycoproteins and their formulations is essential for them to have the desired therapeutic effect on the patient. It has recently been shown that Raman spectroscopy can be used to quantify the proportion of glycosylated protein from mixtures of native and glycosylated forms of bovine pancreatic ribonuclease (RNase). Here we show the first steps toward not only the detection of glycosylation status but the characterization of glycans themselves from just a few protein molecules at a time using tip-enhanced Raman scattering (TERS). While this technique generates complex data that are very dependent on the protein orientation, with the careful development of combined data preprocessing, univariate and multivariate analysis techniques, we have shown that we can distinguish between the native and glycosylated forms of RNase. Many glycoproteins contain populations of subtly different glycoforms; therefore, with stricter orientation control, we believe this has the potential to lead to further glycan characterization using TERS, which would have use in biopharmaceutical synthesis and formulation research

Organic acids, siderophores, enzymes and mechanical pressure for black slate bioweathering with the basidiomycete Schizophyllum commune

Although many fungi are known to be able to perform bioweathering of rocks and minerals, little information is available concerning the role of basidiomycetes in this process. The wood-rotting basidiomycete Schizophyllum commune was investigated for its ability to degrade black slate, a rock rich in organic carbon. Mechanical pressure of hyphae and extracellular polymeric substances was investigated for biophysical weathering. A mixed ß1-3/ß1-6 glucan, likely schizophyllan that is well known from S. commune, could be identified on black slate surfaces. Secretion of siderophores and organic acids as biochemical weathering agents was shown. Both may contribute to biochemical weathering in addition to enzymatic functions. Previously, the exoenzyme laccase was believed to attack organic the matter within the black slate, thereby releasing metals from the rock. Here, overexpression of laccase showed enhanced dissolution of quartz phases by etching and pitting. At the same time, the formation of a new secondary mineral phase, whewellite, could be demonstrated. Hence, a more comprehensive understanding of biophysical as well as biochemical weathering by S. commune could be reached and unexpected mechanisms like quartz dissolution linked to shale degradation. © 2019 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd

Spatially resolved spectroscopic differentiation of hydrophilic and hydrophobic domains on individual insulin amyloid fibrils

The formation of insoluble β-sheet-rich protein structures known as amyloid fibrils is associated with numerous neurodegenerative diseases, such as Alzheimer’s and Parkinson’s disease. A detailed understanding of the molecular structure of the fibril surface is of interest as the first contact with the physiological environment in vivo and plays a decisive role in biological activity and associated toxicity. Recent studies reveal that the inherent sensitivity and specificity of tip-enhanced Raman scattering (TERS) renders this technique a compelling method for fibril surface analysis at the single-particle level. Here, the reproducibility of TERS is demonstrated, indicating its relevance for detecting molecular variations. Consequently, individual fibrils are systematically investigated at nanometer spatial resolution. Spectral parameters were obtained by band-fitting, particularly focusing on the identification of the secondary structure via the amide III band and the differentiation of hydrophobic and hydrophilic domains on the surface. In addition multivariate data analysis, specifically the N-FINDR procedure, was employed to generate structure-specific maps. The ability of TERS to localize specific structural domains on fibril surfaces shows promise to the development of new fibril dissection strategies and can be generally applied to any (bio)chemical surface when structural variations at the nanometer level are of interest

Sphagnum farming on cut-over bog in NW Germany: Long-term studies on Sphagnum growth

Sphagnum farming allows sustainable and climate-friendly land use on bogs while producing a renewable substitute for peat in horticultural growing media. We studied Sphagnum productivity on an experimental Sphagnum culture established on a cut-over bog in Germany with strongly humified peat at the surface. Preparation of the site included levelling of the peat surface, construction of an irrigation system, spreading of Sphagnum papillosum fragments, covering them with straw, and finally rewetting. Provided there was an adequate (95 %) initial cover of Sphagnum fragments, the most relevant variables for Sphagnum productivity were found to be water supply and regular mowing of vascular plants. As long as sufficient water was supplied, the dry biomass accumulation of the established Sphagnum lawn remained high, reaching 3.7 t ha-1 yr-1 between 2007 and 2011. Annual dry Sphagnum biomass productivity over the period 2010–2011 was up to 6.9 t ha-1. During periods when high water table could not be maintained, substantial decomposition of the previously accumulated biomass occurred. After nine years the net accumulated dry mass per hectare was on average 19.5 t of pure Sphagnum and 0.7 t of subsurface vascular-plant biomass. Nitrogen deposition in the study region is apparently sufficient to support fast Sphagnum growth, whereas phosphorus and potassium may be limiting