Highly Porous, Ultralight, Biocompatible Silk Fibroin Aerogel-Based Triboelectric Nanogenerator.

This study presents the fabrication of an ultralight, porous, and high-performance triboelectric nanogenerator (TENG) utilizing silk fibroin (SF) aerogels and PDMS sponges as the friction layer. The transition from two-dimensional film friction layers to three-dimensional porous aerogels significantly increased the specific surface area, offering an effective strategy for designing high-performance SF aerogel-based TENGs. The TENG incorporating the porous SF aerogel exhibited optimal output performance at a 3% SF concentration, achieving a maximum open circuit voltage of 365 V, a maximum short-circuit current of 11.8 µA, and a maximum power density of 7.52 W/m2. In comparison to SF-film-based TENGs, the SF-aerogel based TENG demonstrated a remarkable 6.5-fold increase in voltage and a 4.5-fold increase in current. Furthermore, the power density of our SF-based TENG surpassed the previously reported optimal values for SF-based TENGs by 2.4 times. Leveraging the excellent mechanical stability and biocompatibility of TENGs, we developed an SF-based TENG self-powered sensor for the real-time monitoring of subtle biological movements. The SF-based TENG exhibits promising potential as a wearable bioelectronic device for health monitoring.

Region-selective and site-specific glycation of influenza proteins surrounding the viral envelope membrane.

Analysis of protein modifications is critical for quality control of therapeutic biologics. However, the identification and quantification of naturally occurring glycation of membrane proteins by mass spectrometry remain technically challenging. We used highly sensitive LC MS/MS analyses combined with multiple enzyme digestions to determine low abundance early-stage lysine glycation products of influenza vaccines derived from embryonated chicken eggs and cultured cells. Straightforward sequencing was enhanced by MS/MS fragmentation of small peptides. As a result, we determined a widespread distribution of lysine modifications attributed by the region-selectivity and site-specificity of glycation toward influenza matrix 1, hemagglutinin and neuraminidase. Topological analysis provides insights into the site-specific lysine glycation, localizing in the distinct structural regions of proteins surrounding the viral envelope membrane. Our finding highlights the proteome-wide discovery of lysine glycation of influenza membrane proteins and potential effects on the structural assembly, stability, receptor binding and enzyme activity, demonstrating that the impacts of accumulated glycation on the quality of products can be directly monitored by mass spectrometry-based structural proteomics analyses.

Characterization of Potential Plastic-Degradation Enzymes from Marine Bacteria.

Polyethylene terephthalate (PET) and polyethylene (PE) are prominent polymer materials that comprise a significant portion of commercial plastic waste. Their durability and slow degradation rate have resulted in significant accumulation of plastic on Earth. In a recent study, macrotranscriptomic profiling of a reconstituted marine bacterial community identified 10 putative enzymes capable of directly acting on PE or PET (PEases or PETases). Among these enzymes, three recombinant proteins were reported to possess PE degradation activity. To select potential plastic degrading enzyme candidates for protein engineering efforts, we expressed and purified eight out of the 10 candidates, excluding two due to poor expression and/or solubility. Notably, several candidate proteins displayed significant esterase activity on p-nitrophenyl butyrate and exhibited unexpected thermostability despite their marine origin. Additionally, we observed dose- and time-dependent hydrolytic activity on the PET trimer substrate. Structural analysis and mutagenesis of a candidate protein confirmed the presence of catalytic triad residues, classifying it as an esterase. Furthermore, we elucidated the structural importance of the two disulfide bonds. Through point mutation experiments, we observed an enhanced hydrolytic activity of a selected enzyme candidate on PET nanoparticles. Our findings challenge the classification of the enzymes directly acting on PE and highlight the significance and complexity of validating PE degradation enzymes identified through metagenomic analysis.

Back on the chain gang: polyphosphate modification of proteins.

Polyphosphate (polyP) mediates a plethora of biological functions. Understanding the polyP-protein interactome will help clarify the mechanisms underpinning these functions. Recent studies demonstrating a strong but noncovalent modification of lysine and histidine repeat proteins by polyP have provided new insights into polyP-protein biochemistry with implications for research and therapeutics.

Polyphosphate attachment to lysine repeats is a non-covalent protein modification.

Polyphosphate (polyP) is a chain of inorganic phosphate that is present in all domains of life and affects diverse cellular phenomena, ranging from blood clotting to cancer. A study by Azevedo et al. described a protein modification whereby polyP is attached to lysine residues within polyacidic serine and lysine (PASK) motifs via what the authors claimed to be covalent phosphoramidate bonding. This was based largely on the remarkable ability of the modification to survive extreme denaturing conditions. Our study demonstrates that lysine polyphosphorylation is non-covalent, based on its sensitivity to ionic strength and lysine protonation and absence of phosphoramidate bond formation, as analyzed via 31P NMR. Ionic interaction with lysine residues alone is sufficient for polyP modification, and we present a new list of non-PASK lysine repeat proteins that undergo polyP modification. This work clarifies the biochemistry of polyP-lysine modification, with important implications for both studying and modulating this phenomenon. This Matters Arising paper is in response to Azevedo et al. (2015), published in Molecular Cell. See also the Matters Arising Response by Azevedo et al. (2024), published in this issue.

Analysis of four heavy metal concentrations in sediments fromthe Jiaozhou Bay, China by visible and near infrared spectroscopy (225-975 nm).

As an important component ofbiogeochemical cyclein coastal ecosystems, sediments are the sink of heavy metals. Therefore, distribution and dynamics of heavy metals in sediments could assess ecological quality and predict ecological risks. In the new era, rapid and green technology are highly needed, especially that could determine multi-parameters simultaneously. Here, we explored a new method to rapidly determine concentrations of heavy metals in sediments by visible and near infrared reflectance spectroscopy (VIRS).We sampled sediments in the Jiaozhou Bay, China, collected their reflectance spectra, and measured concentrations of four heavy metals (As, Cr, Cu, and Zn). Heavy metal models were established and evaluated using substances highly correlated with heavy metals. This study provides an effective reference for rapid analysis of As, Cr, Cu, and Zn simultaneously in sediments, at least in the Jiaozhou Bay, and for ecological environment protection and resource development of the Jiaozhou Bay.

Protein purification via consecutive histidine-polyphosphate interaction.

While nickel-nitrilotriacetic acid (Ni-NTA) has greatly advanced recombinant protein purification, its limitations, including nonspecific binding and partial purification for certain proteins, highlight the necessity for additional purification such as size exclusion and ion exchange chromatography. However, specialized equipment such as FPLC is typically needed but not often available in many laboratories. Here, we show a novel method utilizing polyphosphate (polyP) for purifying proteins with histidine repeats via non-covalent interactions. Our study demonstrates that immobilized polyP efficiently binds to histidine-tagged proteins across a pH range of 5.5-7.5, maintaining binding efficacy even in the presence of reducing agent DTT and chelating agent EDTA. We carried out experiments of purifying various proteins from cell lysates and fractions post-Ni-NTA. Our results demonstrate that polyP resin is capable of further purification post-Ni-NTA without the need for specialized equipment and without compromising protein activity. This cost-effective and convenient method offers a viable approach as a complementary approach to Ni-NTA.

Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli.

Polyphosphate exhibits a unique post-translational modification-like function, known as histidine polyphosphate modification (HPM), marked by a robust non-covalent interaction with histidine repeat proteins. Here, we present a protocol for detecting HPM of human proteins via maltose-binding protein-tagged expression in E. coli. We describe steps for detecting HPM by observing electrophoretic mobility shifts on NuPAGE gels followed by western blot. We then detail procedures for analyzing the influence of ionic strength and pH on HPM. For complete details on the use and execution of this protocol, please refer to Neville et al.1.