Solid-state NMR spectroscopy of functional amyloid from a fungal hydrophobin: A well-ordered β-sheet core amidst structural heterogeneity.

GrEASy fibrils: Hydrophobins are fungal proteins that assemble into an amphipathic fibrillar monolayer with amyloid properties and a hydrophobic face as water-resistant as Teflon. Solid-state NMR studies on EAS hydrophobin fibrils reveal direct evidence of a partial molecular rearrangement on assembly and an ordered β-sheet-rich core in the context of a whole protein in this functional amyloid

Non-equilibrium hydrogen exchange for determination of H-bond strength and water accessibility in solid proteins.

We demonstrate measurement of non-equilibrium backbone amide hydrogen-deuterium exchange rates (HDX) for solid proteins. The target of this study are the slowly exchanging residues in solid samples, which are associated with stable secondary-structural elements of proteins. These hydrogen exchange processes escape methods measuring equilibrium exchange rates of faster processes. The method was applied to a micro-crystalline preparation of the SH3 domain of chicken α-spectrin. Therefore, from a 100% back-exchanged micro-crystalline protein preparation, the supernatant buffer was exchanged by a partially deuterated buffer to reach a final protonation level of approximately 20% before packing the sample in a 1.3 mm rotor. Tracking of the HN peak intensities for 2 weeks reports on site-specific hydrogen bond strength and also likely reflects water accessibility in a qualitative manner. H/D exchange can be directly determined for hydrogen-bonded amides using 1H detection under fast magic angle spinning. This approach complements existing methods and provides the means to elucidate interesting site-specific characteristics for protein functionality in the solid state

Characterization of soil organic matter in aggregates and size-density fractions by solid state C-13 CPMAS NMR spectroscopy.

Understanding the changes in soil organic matter (SOM) composition during aggregate formation is crucial to explain the stabilization of SOM in aggregates. The objectives of this study were to investigate (i) the composition of SOM associated with different aggregates and size-density fractions and (ii) the role of selective preservation in determining the composition of organic matter in aggregate and size-density fractions. Surface soil samples were collected from an Alfisol on the Northern Tablelands of NSW, Australia with contrasting land uses native pasture, crop-pasture rotation and woodland. Solid state 13C cross-polarization and magic angle spinning (CPMAS) Nuclear Magnetic Resonance (NMR) spectroscopy was used to determine the SOM composition in macroaggregates (250-2000 µm), microaggregates (53-250 µm), and <53 µm fraction. The chemical composition of light fraction (LF), coarse particulate organic matter (cPOM), fine particulate organic matter (fPOM) and mineral associated soil organic matter (mSOM) were also determined. The major constituent of SOM of aggregate size fractions was O-alkyl carbon, which represented 44-57% of the total signal acquired, whereas alkyl carbon contributed 16-27%. There was a progressive increase in alkyl carbon content with decrease in aggregate size. Results suggest that SOM associated with <53 µm fraction was at a more advanced stage of decomposition than that of macroaggregates and microaggregates. The LF and cPOM were dominated by O-alkyl carbon while alkyl carbon content was high in fPOM and mSOM. Interestingly, the relative change in O-alkyl, alkyl and aromatic carbon between aggregates and SOM fractions revealed that microbial synthesis and decomposition of organic matter along with selective preservation of alkyl and aromatic carbon plays a significant role in determining the composition of organic matter in aggregates

Access to side-chain carbon information in deuterated solids under fast MAS through non-rotor-synchronized mixing.

We demonstrate the accessibility of aliphatic 13C side chain chemical shift sets for solid-state NMR despite perdeuteration and fast MAS using isotropic, non-rotor-synchronized 13C-13C mixing. Combined with amide proton detection, we unambiguously and sensitively detect whole side chain to backbone correlations for two proteins using around 1 mg of sample

A disilene base adduct with a dative Si–Si single bond.

An experimental and theoretical study of the base- stabilized disilene 1 is reported, whichforms at lowtemper- atures in the disproportionation reaction of Si 2 Cl 6 or neo- Si 5 Cl 12 with equimolar amounts of NMe 2 Et. Single-crystal X- ray diffraction and quantum-chemical bonding analysis dis- close an unprecedented structure in silicon chemistry featuring adative Si!Si single bond between two silylene moieties, Me 2 EtN!SiCl 2 !Si(SiCl 3 ) 2 .The central ambiphilic SiCl 2 group is linked by dative bonds to the amine donor and the bis(trichlorosilyl)silylene acceptor,which leads to push–pull stabilization. Based on experimental and theoretical examina- tions aformation mechanism is presented that involves an autocatalytic reaction of the intermediately formed anion Si(SiCl 3 ) 3 ¢ with neo-Si 5 Cl 12 to yield 1

Impairment of germline transmission after blastocyst injection with murine embryonic stem cells cultured with mouse hepatitis virus and mouse minute virus

The aim of this study was to determine the susceptibility of murine embryonic stem (mESCs) to mouse hepatitis virus (MHV-A59) and mouse minute virus (MMVp) and the effect of these viruses on germline transmission (GLT) and the serological status of recipients and pups. When recipients received 10 blastocysts, each injected with 100 TCID50 MHV-A59, three out of five recipients and four out of 14 pups from three litters became seropositive. When blastocysts were injected with 10−5 TCID50 MMVp, all four recipients and 14 pups from four litters remained seronegative. The mESCs replicated MHV-A59 but not MMVp, MHV-A59 being cytolytic for mESCs. Exposure of mESCs to the viruses over four to five passages but not for 6 h affected GLT. Recipients were seropositive for MHV-A59 but not for MMVp when mESCs were cultured with the virus over four or five passages. The data show that GLT is affected by virus-contaminated mESCs

Internet of Things for Sustainable Forestry

Forests and grasslands play an important role in water and air purification, prevention of the soil erosion, and in provision of habitat to wildlife. Internet of Things has a tremendous potential to play a vital role in the forest ecosystem management and stability. The conservation of species and habitats, timber production, prevention of forest soil degradation, forest fire prediction, mitigation, and control can be attained through forest management using Internet of Things. The use and adoption of IoT in forest ecosystem management is challenging due to many factors. Vast geographical areas and limited resources in terms of budget and equipment are some of the limiting factors. In digital forestry, IoT deployment offers effective operations, control, and forecasts for soil erosion, fires, and undesirable depositions. In this chapter, IoT sensing and communication applications are presented for digital forestry systems. Different IoT systems for digital forest monitoring applications are also discussed

Fractional deuteration applied to biomolecular solid-state NMR spectroscopy

Solid-state Nuclear Magnetic Resonance can provide detailed insight into structural and dynamical aspects of complex biomolecules. With increasing molecular size, advanced approaches for spectral simplification and the detection of medium to long-range contacts become of critical relevance. We have analyzed the protonation pattern of a membrane-embedded ion channel that was obtained from bacterial expression using protonated precursors and D2O medium. We find an overall reduction of 50% in protein protonation. High levels of deuteration at Hα and Hβ positions reduce spectral congestion in (1H,13C,15N) correlation experiments and generate a transfer profile in longitudinal mixing schemes that can be tuned to specific resonance frequencies. At the same time, residual protons are predominantly found at amino-acid side-chain positions enhancing the prospects for obtaining side-chain resonance assignments and for detecting medium to long-range contacts. Fractional deuteration thus provides a powerful means to aid the structural analysis of complex biomolecules by solid-state NMR

Structural Analysis of a Peptide Fragment of Transmembrane Transporter Protein Bilitranslocase

Using a combination of genomic and post-genomic approaches is rapidly altering the number of identified human influx carriers. A transmembrane protein bilitranslocase (TCDB 2.A.65) has long attracted attention because of its function as an organic anion carrier. It has also been identified as a potential membrane transporter for cellular uptake of several drugs and due to its implication in drug uptake, it is extremely important to advance the knowledge about its structure. However, at present, only the primary structure of bilitranslocase is known. In our work, transmembrane subunits of bilitranslocase were predicted by a previously developed chemometrics model and the stability of these polypeptide chains were studied by molecular dynamics (MD) simulation. Furthermore, sodium dodecyl sulfate (SDS) micelles were used as a model of cell membrane and herein we present a high-resolution 3D structure of an 18 amino acid residues long peptide corresponding to the third transmembrane part of bilitranslocase obtained by use of multidimensional NMR spectroscopy. It has been experimentally confirmed that one of the transmembrane segments of bilitranslocase has alpha helical structure with hydrophilic amino acid residues oriented towards one side, thus capable of forming a channel in the membrane