Mechanistic Insight into the Reactivation of BCAII Enzyme from Denatured and Molten Globule States by Eukaryotic Ribosomes and Domain V rRNAs

In all life forms, decoding of messenger-RNA into polypeptide chain is accomplished by the ribosome. Several protein chaperones are known to bind at the exit of ribosomal tunnel to ensure proper folding of the nascent chain by inhibiting their premature folding in the densely crowded environment of the cell. However, accumulating evidence suggests that ribosome may play a chaperone role in protein folding events in vitro. Ribosome-mediated folding of denatured proteins by prokaryotic ribosomes has been studied extensively. The RNA-assisted chaperone activity of the prokaryotic ribosome has been attributed to the domain V, a span of 23S rRNA at the intersubunit side of the large subunit encompassing the Peptidyl Transferase Centre. Evidently, this functional property of ribosome is unrelated to the nascent chain protein folding at the exit of the ribosomal tunnel. Here, we seek to scrutinize whether this unique function is conserved in a primitive kinetoplastid group of eukaryotic species Leishmania donovani where the ribosome structure possesses distinct additional features and appears markedly different compared to other higher eukaryotic ribosomes. Bovine Carbonic Anhydrase II (BCAII) enzyme was considered as the model protein. Our results manifest that domain V of the large subunit rRNA of Leishmania ribosomes preserves chaperone activity suggesting that ribosome-mediated protein folding is, indeed, a conserved phenomenon. Further, we aimed to investigate the mechanism underpinning the ribosome-assisted protein reactivation process. Interestingly, the surface plasmon resonance binding analyses exhibit that rRNA guides productive folding by directly interacting with molten globule-like states of the protein. In contrast, native protein shows no notable affinity to the rRNA. Thus, our study not only confirms conserved, RNA-mediated chaperoning role of ribosome but also provides crucial insight into the mechanism of the process

Effect of particle and carbide grain sizes on a HVOAF WC-Co-Cr coating for the future application on internal surfaces: microstructure and wear

The use of nanoscale WC grain or finer feedstock particles are possible methods of improving the performance of WC-Co-Cr coatings. Finer powders are being pursued for the development of coating internal surfaces, as less thermal energy is required to melt the finer powder compared to coarse powders, permitting spraying at smaller stand - off distances. Three WC-0Co-4Cr coatings, with two different powder particle sizes and two different carbide grain sizes, were sprayed using a high velocity oxy-air fuel (HVOAF) thermal spray system developed by Castolin Eutectic - Monitor Coatings Ltd., UK. Powder and coating microstructures were characterised using XRD and SEM. Fracture toughness and dry sliding wear performance at three loads were investigated using a ball–on-disc tribometer with a WC–Co counter body. It was found that the finer powder produced the coating with the highest microhardness, but its fracture toughness was reduced due to increased decarburisation compared to the other powders. The sprayed nanostructured powder had the lowest microhardness and fracture toughness of all materials tested. Unlubricated sliding wear testing at the lowest load showed the nanostructured coating performed best; however at the highest load this coating showed the highest specific wear rates with the other two powders performing to a similar, better standard

Structure–function relationships in fluorescent marker proteins of the green fluorescent protein family

GFP-like proteins, originally cloned from marine animals, are genetically encoded fluorescence markers that have become indispensable tools for the life sciences. The search for GFP-like proteins with novel and improved properties is ongoing, driven by the persistent need for advanced and specialized fluorescence labels for cellular imaging. The 3D structures of these proteins are overall similar. However, considerable variations have been found in the covalent structures and the stereochemistry of the chromophore, which govern essential optical properties such as the absorption/emission wavelengths. A detailed understanding of the structure and dynamics of GFP-like proteins greatly aids in the rational engineering of advanced fluorescence marker proteins. In this chapter, we summarize the present knowledge of the structural diversity of GFP-like proteins and discuss how structure and dynamics govern their optical properties

Advances in Corrosion-Resistant Thermal Spray Coatings for Renewable Energy Power Plants : Part II - Effect of Environment and Outlook

High-temperature corrosion of critical components such as water walls and superheater tubes in biomass/waste-fired boilers is a major challenge. A dense and defect-free thermal spray coating has been shown to be promising to achieve a high electrical/thermal efficiency in power plants. The field of thermal spraying and quality of coatings have been progressively evolving; therefore, a critical assessment of our understanding of the efficacy of coatings in increasingly aggressive operating environments of the power plants can be highly educative. The effects of composition and microstructure on high-temperature corrosion behavior of the coatings were discussed in the first part of the review. The present paper that is the second part of the review covers the emerging research field of performance assessment of thermal spray coatings in harsh corrosion-prone environments and provides a comprehensive overview of the underlying high-temperature corrosion mechanisms that lead to the damage of exposed coatings. The application of contemporary analytical methods for better understanding of the behavior of corrosion-resistant coatings is also discussed. A discussion based on an exhaustive review of the literature provides an unbiased commentary on the advanced accomplishments and some outstanding issues in the field that warrant further research. An assessment of the current status of the field, the gaps in the scientific understanding, and the research needs for the expansion of thermal spray coatings for high-temperature corrosion applications is also provided. © 2019, The Author(s)