Structure of the human galanin receptor 2 bound to galanin and Gq reveals the basis of ligand specificity and how binding affects the G-protein interface.

Galanin is a neuropeptide expressed in the central and peripheral nervous systems, where it regulates various processes including neuroendocrine release, cognition, and nerve regeneration. Three G-protein coupled receptors (GPCRs) for galanin have been discovered, which is the focus of efforts to treat diseases including Alzheimer's disease, anxiety, and addiction. To understand the basis of the ligand preferences of the receptors and to assist structure-based drug design, we used cryo-electron microscopy (cryo-EM) to solve the molecular structure of GALR2 bound to galanin and a cognate heterotrimeric G-protein, providing a molecular view of the neuropeptide binding site. Mutant proteins were assayed to help reveal the basis of ligand specificity, and structural comparison between the activated GALR2 and inactive hß2AR was used to relate galanin binding to the movements of transmembrane (TM) helices and the G-protein interface.

Structural and Functional Characterizations of Cancer Targeting Nanoparticles Based on Hepatitis B Virus Capsid.

Cancer targeting nanoparticles have been extensively studied, but stable and applicable agents have yet to be developed. Here, we report stable nanoparticles based on hepatitis B core antigen (HBcAg) for cancer therapy. HBcAg monomers assemble into spherical capsids of 180 or 240 subunits. HBcAg was engineered to present an affibody for binding to human epidermal growth factor receptor 1 (EGFR) and to present histidine and tyrosine tags for binding to gold ions. The HBcAg engineered to present affibody and tags (HAF) bound specifically to EGFR and exterminated the EGFR-overexpressing adenocarcinomas under alternating magnetic field (AMF) after binding with gold ions. Using cryogenic electron microscopy (cryo-EM), we obtained the molecular structures of recombinant HAF and found that the overall structure of HAF was the same as that of HBcAg, except with the affibody on the spike. Therefore, HAF is viable for cancer therapy with the advantage of maintaining a stable capsid form. If the affibody in HAF is replaced with a specific sequence to bind to another targetable disease protein, the nanoparticles can be used for drug development over a wide spectrum.

The effect of blood cadmium levels on hypertension in male firefighters in a metropolitan city.

Background: This study investigated the effect of dispatch frequency on blood cadmium levels and the effect of blood cadmium levels on hypertension in male firefighters in a metropolitan city. Methods: We conducted a retrospective longitudinal study of male firefighters who completed the regular health checkups, including a health examination survey and blood cadmium measurements. We followed them for 3 years. To investigate the effect of dispatch frequency on blood cadmium levels and the effect of blood cadmium levels on hypertension, we estimated the short-term (model 1) and long-term (model 2) effects of exposure and hypothesized a reversed causal pathway model (model 3) for sensitivity analysis. Sequential conditional mean models were fitted using generalized estimating equations, and the odds ratios (ORs) and the respective 95% confidence intervals (CIs) were calculated for hypertension for log-transformed (base 2) blood cadmium levels and quartiles. Results: Using the lowest category of dispatch frequency as a reference, we observed that the highest category showed an increase in blood cadmium levels of 1.879 (95% CI: 0.673, 3.086) µg/dL and 0.708 (95% CI: 0.023, 1.394) µg/dL in models 2 and 3, respectively. In addition, we observed that doubling the blood cadmium level significantly increased the odds of hypertension in model 1 (OR: 1.772; 95% CI: 1.046, 3.003) and model 3 (OR: 4.288; 95% CI: 1.110, 16.554). Using the lowest quartile of blood cadmium levels as a reference, the highest quartile showed increased odds of hypertension in model 1 (OR: 2.968; 95% CI: 1.121, 7.861) and model 3 (OR: 33.468; 95% CI: 1.881, 595.500). Conclusions: We found that dispatch frequency may affect blood cadmium levels in male firefighters, and high blood cadmium levels may influence hypertension in a dose-response manner.

Cryo-EM structure of the human somatostatin receptor 2 complex with its agonist somatostatin delineates the ligand-binding specificity.

Somatostatin is a peptide hormone that regulates endocrine systems by binding to G-protein-coupled somatostatin receptors. Somatostatin receptor 2 (SSTR2) is a human somatostatin receptor and is highly implicated in hormone disorders, cancers, and neurological diseases. Here, we report the high-resolution cryo-EM structure of full-length human SSTR2 bound to the agonist somatostatin (SST-14) in complex with inhibitory G (Gi) proteins. Our structural and mutagenesis analyses show that seven transmembrane helices form a deep pocket for ligand binding and that SSTR2 recognizes the highly conserved Trp-Lys motif of SST-14 at the bottom of the pocket. Furthermore, our sequence analysis combined with AlphaFold modeled structures of other SSTR isoforms provide a structural basis for the mechanism by which SSTR family proteins specifically interact with their cognate ligands. This work provides the first glimpse into the molecular recognition mechanism of somatostatin receptors and a crucial resource to develop therapeutics targeting somatostatin receptors.

Structural and functional identification of the uncharacterized metallo-ß-lactamase superfamily protein TW9814 as a phosphodiesterase with unique metal coordination.

Metallo-ß-lactamase (MBL) superfamily proteins have a common αß/ßα sandwich fold and perform a variety of functions through metal-mediated catalysis. However, because of the enormous scale of this superfamily, only a small percentage of the proteins belonging to the superfamily have been annotated structurally or functionally to date. Therefore, much remains unknown about the MBL superfamily proteins. Here, TW9814, a hypothetical MBL superfamily protein, was structurally and functionally investigated. Guided by the crystal structure of dimeric TW9814, it was demonstrated that TW9814 functions as a phosphodiesterase (PDE) in the presence of divalent metal ions such as manganese(II) or nickel(II). A docking model between TW9814 and the substrate bis(p-nitrophenyl)phosphate (bpNPP) showed the importance of the dimerization of TW9814 for its bpNPP-hydrolyzing activity and for the interaction between the enzyme and the substrate. TW9814 showed outstanding catalytic efficiency (kcat/Km) under alkaline conditions compared with other PDEs. The activity of TW9814 appears to be regulated through a disulfide bond, which is a feature that is not present in other MBL superfamily members. This study provides a platform for the functional characterization of other hypothetical proteins of the MBL or other superfamilies.

In-Bi Electrocatalyst for the Reduction of CO2 to Formate in a Wide Potential Window.

The CO2 atmospheric concentration level hit the record at more than 400 ppm and is predicted to keep increasing as the dependence on fossil fuels is inevitable. The CO2 electrocatalytic conversion becomes an alternative due to its environmental and energy-friendly properties and benign operation condition. Lately, bimetallic materials have drawn significant interest as electrocatalysts due to their distinct properties, which the parents' metal cannot mimic. Herein, the indium-bismuth nanosphere (In16Bi84 NS) was fabricated via the facile liquid-polyol technique. The In16Bi84 NS exhibits exceptional performance for CO2 reduction to formate, with the faradaic efficiency (FE) approaching ∼100% and a corresponding partial current density of 14.1 mA cm-2 at -0.94 V [vs the reversible hydrogen electrode (RHE)]. Furthermore, the FE could be maintained above 90% in a wide potential window (-0.84 to -1.54 V vs the RHE). This superior performance is attributed to the tuned electronic properties induced by the synergistic interaction between In and Bi, enabling the intermediates to be stably adsorbed on the catalyst surface to generate more formate ions.

Related factors for preserving firefighter's pulmonary function.

BACKGROUND: Firefighters are constantly exposed to harmful substances in the respiratory tract and require management measures. We comprehensively compared factors affecting the lung function of firefighters to identify management measures that can reduce the deterioration of lung function. METHODS: A cross-sectional study was conducted in 1,108 male firefighters. Subjects were surveyed with self-written questionnaires that included a history of smoking, number of workouts per week, work department, and medical history, including diseases that could affect lung function. Body mass index was calculated using an automatic body measurement instrument and body fat, body fat percentage, muscle mass, and skeletal muscle mass were measured using Inbody 770. Based on the body weight obtained from body measurements, skeletal muscle mass height-adjusted skeletal muscle index (hSMI) compared to height was determined. For lung function, forced expiratory volume in one second (FEV1), forced vital capacity (FVC), FEV1/FVC, and peak expiratory flow (PEF) were measured using a spirometer HI-801. Analysis of variance and independent t-tests were performed for univariate analysis of factors that could affect lung function, and multiple regression analysis was performed for multivariate analysis. RESULTS: When the factors relating lung function were analyzed using regression analysis, FEV1 was negatively correlated with age, body fat percentage, and duty year, positively with height and hSMI. FVC increased with height and hSMI, decreased with age, body fat percentage, and duty year. FEV1/FVC was related with age, height, body fat percentage and working history. Height and muscle mass were analyzed as related factors on PEF. When the analysis was conducted on firefighters who exercised more than 3 times a week, working history had lower relation with FEV1 and body fat percentage had no relation with FEV1/FVC. CONCLUSION: We suggest management measures to reduce body fat percentage and increase skeletal muscle mass to maintain lung function in firefighters.