Enzymes are pivotal regulators of cellular metabolism and organismal homeostasis, and their dysregulation is often associated with the onset and progression of disease. Therefore, accurate monitoring of the real activities of target enzymes is essential for deciphering biological mechanisms and gaining pathological insight. Through decades of iterative refinement, a diverse repertoire of enzyme-activatable fluorogenic probes (EAFPs) have been developed, enabling the capture of aberrant enzyme dynamics with high spatiotemporal resolution and multifunctional biosensing capabilities. In this context, we highlight the current state-of-the-art EAFPs, spanning fundamental design principles to proof-of-concept applications. First, the molecular engineering, sensing mechanisms, and design strategies of EAFPs are introduced. Next, a wide range of cutting-edge probes for imaging and sensing target enzyme(s) are presented, with emphasis on structural features, recognition mechanisms, and biomedical applications. Representative examples in biomarker imaging, disease diagnosis, drug screening, and therapeutic testing are highlighted to illustrate both the design principles and practical utility. Finally, the existing challenges and future trajectories for EAFPs in specific application scenarios are discussed. The insights presented here will inspire and accelerate the development of high-performance multifunctional EAFPs for both fundamental and translational research.
Protein folding is essential for maturation of nascent polypeptides into native proteins. In living cells, molecular chaperones and enzymes efficiently and synergistically promote protein folding. Inspired by biological folding systems, researchers have developed synthetic chemical chaperones for promoting protein folding in vitro. Many secretory proteins have multiple disulfide (SS) bonds, and therefore, their folding reactions are intrinsically an oxidative process, namely, oxidative folding. This review highlights representative strategies in the design of redox-active compounds that promote oxidative protein folding. Small-molecule thiols and disulfides were developed for oxidative folding promotion, which have been improved by optimizing their molecular structures and reactivities. The chemical features of natural folding enzymes have also inspired researchers to develop folding promoters that reproduce the structural and chemical features of the enzymes. Biological contexts have been considered recently, giving birth to stress-responsive folding promoters. These efforts described herein would pave the way for the development of novel synthetic reagents that enhance industrial protein production processes and prevent protein misfolding diseases.
The escalating environmental impact of polyethylene terephthalate (PET) waste, particularly from synthetic fabrics, necessitates sustainable degradation strategies. This study focused on the screening of potent fungal isolates capable of degrading PET, with a specific emphasis on optimizing lipase production, a key enzyme involved in ester bond hydrolysis. The isolates were primarily screened for the production of PET degradation enzymes qualitatively and quantitatively. Among the screened isolates, PPS3 showed the highest PET fabric waste degradation efficiency of 13.6 ± 1.31% in quantitative screening. Based on 18S rRNA partial gene sequencing, this potent isolate was identified as Aspergillus niger and was utilized for further optimization and degradation experiments. Bioprocess variables influencing fungal lipase enzyme production were optimized through 'One-factor-at-a-time' (OFAT) approach. The optimized lipase production media was inoculated with the potent fungal isolate along with the PET waste and kept for 10 weeks of incubation. A weight loss of approximately 55 ± 2.38% was recorded, corresponding to the reduction in PET substrate mass, which demonstrates the degradation potential of the lipase enzyme. The PET degradation potential of the selected isolate was assessed through surface morphology changes (FESEM), and functional group modification (FTIR). Results demonstrated substantial PET fabric degradation, confirming the efficacy of the fungal strain and the importance of lipase in the biodegradation process. This study presents a promising biotechnological approach for mitigating PET pollution through microbial intervention and enzyme optimization.
Using atomic force microscopy (AFM), the heights of individual horseradish peroxidase (HRP) molecules were determined at different stages of the catalytic cycle of this enzyme (I-IV). The HRP heights were determined in two buffer systems: Dulbecco's phosphate-buffered saline (PBSD) (pH 7.2) and citrate-phosphate buffer (CPB) (pH 5.0). Spectrophotometric analysis showed that the enzyme was more active in CPB. AFM data processing under these conditions revealed a tendency toward a decrease in the HRP molecule height after addition of the ABTS substrate (stage (II)). After the addition of H2O2 (stage (III)), an increase in the height was observed followed by a further decrease after the addition of the NaN3 inhibitor (stage (IV)). No such trend was observed in PBSD. The increase in the molecular height at stage (III) (in the presence of all enzyme system components) was interpreted as an evidence of the HRP activity. The proportion of molecules with the increased height at stage (III) reached 50.5% in CPB and did not exceed 14.8% in PBSD. These data support the existence of "active" and "non-active" HRP molecules in the studied sample, thus confirming the need to move from traditional biochemical "enzyme activity" to identifying "active" and "non-active" enzyme molecules using a fundamentally new approach of analysis of the properties of individual molecules.
This study investigated the molecular mechanism underlying berberine (BER)-mediated inhibition of acetylcholinesterase (AChE) and its synergistic interaction with galantamine hydrobromide (GAL). Heatmap-based dose-response analysis revealed significant synergistic inhibitory effect of BER and GAL on AChE activity at concentrations of 4.0 μM BER and 8.0 μM GAL. Enzyme kinetic analyses demonstrated that GAL, BER, and the BER GAL combination exhibited competitive, non-competitive, and mixed-type inhibition modes against AChE, respectively. Fluorescence quenching and circular dichroism spectroscopy confirmed high affinity binding of BER to AChE, predominantly stabilized by hydrophobic interactions and hydrogen bonding resulting in conformational alterations in the enzyme structure. Molecular docking simulations further indicated that BER binding affects the solvent-accessible surface area of AChE, induces microenvironmental changes within AChE, and enhances its hydrophobicity. In addition, molecular docking simulations confirmed that BER bound to the peripheral anionic site (PAS) (Trp286, Tyr341, Tyr124) of AChE inducing a more flexible enzyme conformation that facilitates more sites of BER binding. This structural loosening hinders substrate access to the catalytic active site (CAS), thereby inhibiting AChE's catalytic activity. BER mediated PAS remodeling, which enhances GAL's binding affinity for the CAS and cooperatively destabilizes the AChE structure ultimately disrupting substrate-enzyme interactions.
Protein and nucleic acid alkylation are important genetic and epigenetic modifications. The dynamic balance between alkylation and dealkylation is regulated by distinct sets of enzymes and is essential for maintaining genomic stability. Escherichia coli AlkB is a member of the Alkylation B (AlkB) family of dioxygenases that dealkylates a wide range of nucleic acid substrates in E. coli, thereby playing a crucial role in cellular repair processes and epigenetic regulation. Here, we report the backbone 1H, 15N, 13C chemical shift assignment of E. coli AlkB in complex with Zn2+ and α-ketoglutarate. Experiments were acquired at 20 °C by heteronuclear multidimensional NMR spectroscopy. Collectively, 91% of all 13C, 15N and 1H backbone resonances of Alkb were assigned, with 182 out of a possible 200 residues assigned in the 1H-15N TROSY spectrum. Using the program TALOS + , a secondary structure prediction was generated from the assigned backbone resonances that is consistent with the previously reported X-ray structure of the enzyme. The reported assignment will permit investigations of the protein structural dynamics anticipated to provide crucial insight regarding fundamental aspects in the recognition and enzyme regulation processes.
A novel series of 1,2,4-triazole-benzoxazepine hybrid derivatives was designed, synthesized, and biologically evaluated as α-glucosidase inhibitors using an in vitro enzyme inhibition assay with acarbose as a reference standard. Several compounds demonstrated significant inhibitory activity, with compound 14 emerging as the most potent inhibitor (IC₅₀ = 49.26 µM), surpassing acarbose by approximately 3.6-fold, while compound 28 also showed notable activity (IC₅₀ = 72.14 µM). Structure-activity relationship analysis revealed that halogen substitution, particularly at the ortho position, plays a critical role in enhancing inhibitory potency. Molecular docking studies (PDB ID: 7KB6) provided mechanistic insights into ligand-enzyme interactions, showing that the most active compounds establish key π-π stacking interactions with residues TRP423, TRP525, and PHE307, along with π-anion interactions involving ASP640 and ASP564, and extensive hydrophobic contacts with PHE571, PHE673, MET565, and ARG624. Additionally, water-mediated hydrogen bonding (e.g., HOH1356, HOH1625) helps stabilize the ligand within the active site. These interactions enable simultaneous engagement of the catalytic active site and peripheral regions, supporting strong binding affinity. Overall, these results define clear structure-activity relationships and identify 1,2,4-triazole-benzoxazepine hybrids as promising lead scaffolds for further optimization toward novel antidiabetic agents.
A series of chalcone derivatives (8a-8v) was designed and synthesized, and their inhibitory activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were evaluated. The inhibitory activities of AChE and BuChE were measured using the Ellman method. The results showed that most compounds exhibited moderate to weak inhibitory activity against both AChE and BuChE. Among them, compound 8g displayed potent AChE inhibitory activity with an IC50 of 2.59 μM and moderate BuChE inhibitory activity with an IC50 of 8.89 μM. The inhibitory effects of compound 8g on both enzymes surpassed those of the positive control, galantamine. Next, the antioxidant activity of these compounds was assessed using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay, revealing that compound 8j had the strongest antioxidant activity, with an IC50 of 65.84 μM, while the others showed weak antioxidant activity. Enzyme kinetic studies confirmed that compound 8g acts as a mixed-type inhibitor. Molecular docking indicated that compound 8g interacts with both the catalytic active site (CAS) and peripheral anion site (PAS) of AChE. Molecular dynamics (MD) simulations verified the stability of the 8g-AChE/BuChE complex. Overall, these experimental results suggest that the designed and synthesized cholinesterase inhibitor (ChEI) 8g has potential for further development.
Rhodoquinone (RQ) is a crucial electron carrier involved in anaerobic metabolism across select bacteria, protists, and animal species. Its biosynthesis is catalyzed by the rhodoquinone biosynthesis enzyme (RquA), a methyltransferase-like enzyme that uses S-adenosyl-L-methionine to transfer an amino group, converting ubiquinone (UQ) into RQ. The activity of RquA in vitro is enhanced by the presence of divalent metal cations. To probe the metal dependence of RquA, we characterized its interactions with Mn(II), Co(II), and Zn(II). We found that these metal cations bind to RquA with a 1:1 stoichiometry and that Mn(II) and Co(II) exhibited sub-micromolar binding affinities to RquA. Using Mn(II) as a spectroscopic probe, continuous‑wave electron paramagnetic resonance (EPR) revealed a single Mn(II) species with zero‑field splitting parameters |D| = 540(30) MHz and E/D = 0.30(3). Pulse EPR experiments on natural‑abundance and 15N‑labeled samples further identified a weakly‑to‑moderately coupled nitrogen ligand, with an isotropic hyperfine coupling constant |aiso(15N)| = 2.7(1) MHz. Integrating these data with an AlphaFold3‑derived structural model, we propose a putative binding site for the catalytically required divalent metal cation, providing new insight into the structural basis of RquA function.
Single-target ligands have insufficient effectiveness in treating Alzheimer's disease (AD), leading to the development of new pharmacological strategies, particularly multi-target-directed ligands (MTDLs) that tackle the multifactorial nature of the impairment. Among these, dual inhibition of acetylcholinesterase (AChE) and monoamine oxidase B (MAO-B) represents a promising approach for enhancing therapeutic outcomes. Here, analogs of 4-methylbenzyl 5-arylthiophene-2-carboxylates (5a-5h) were identified as dual inhibitors of AChE and MAO-B. In vitro evaluations demonstrated that 5a-5d exhibited the most promising inhibition potential towards targeted enzymes (AChE and MAO-B), having IC50 values 0.72 ± 0.01 µM to 1.69 ± 0.04 µM for AChE and 0.19 ± 0.03 µM to 2.69 ± 0.10 µM for MAO-B. Docking analysis is consistent with the in vitro studies, critically unveiling bindings, commonly hydrogen interactions, π-Sulphur, π-π interaction, π-alkyl, and alkyl-alkyl ligand and enzyme binding interactions. These results underscore the promise of these dual inhibitors in tackling the complex pathology of AD.
The aim of this study was to synthesize Tetrahydroisoquinoline Derivative 1 (M1) and to evaluate its biological activities in the SH-SY5Y neuroblastoma cell line. Theoretical calculations for the investigated molecule were performed using the Gaussian software package at the B3LYP, HF, and M062X levels with the 6-31g, 6-31++g, and 6-31++g(d,p) basis sets. Subsequently, the activity of the compound against SH-SY5Y cancer-related proteins (PDB IDs: 2F37, 3PBL, and 5WIV) was assessed. In addition, the molecule-likeness properties of the molecule were evaluated through ADME/T analyses. The cytotoxic activity of M1 in the SH-SY5Y cell lines was determined using the MTT assay. Following treatment with M1, the expression levels of apoptosis-related genes (MYC, CASP2, BAX, and NF-κB1) and genes associated with DNA repair mechanisms (TP53, RAD51, BRCA2, and MDM2) were analyzed by RT-PCR. Enzyme activities were also measured in M1-treated SH-SY5Y cells. The results demonstrated that M1 exerted its highest cytotoxic effect in the SH-SY5Y cell line after 72 h of incubation. Compared with the control group, M1 showed a stronger effect on G6PDH activity in SH-SY5Y cells, while catalase activity increased by 78% following M1 treatment. Moreover, M1 markedly reduced cell viability in SH-SY5Y cells relative to the control group. In conclusion, these findings indicate that Tetrahydroisoquinoline Derivative M1 exhibits pronounced cytotoxic activity in SH-SY5Y neuroblastoma cells and significantly modulates oxidative stress-related enzyme activities as well as the expression of genes involved in apoptosis and DNA repair pathways, suggesting that M1 may represent a novel and promising candidate for neuroblastoma therapy.
This study evaluated the effects of genotype, dietary ginger supplementation, and genotype × treatment interactions on growth performance, serum biochemical indices, liver and kidney function markers, and oxidative status of broiler chickens. A total of 200 day-old broiler chicks comprising 100 Abor Acres and 100 Cobb 500 were randomly allotted to four dietary treatments containing 0, 0.5, 1.0, and 1.5% ginger powder in a 2 × 4 factorial arrangement within a completely randomized design. Each treatment consisted of 24 birds with three replicates of eight birds, and the trial lasted 56 days. Genotype significantly affected performance, renal markers, and antioxidant status (p ≤ 0.05), with Cobb 500 generally exhibiting superior values, whereas Abor Acres showed significantly higher serum biochemical and liver enzyme values. Dietary ginger supplementation also significantly influenced performance, serum biochemical parameters, renal markers, and antioxidant status (p ≤ 0.05). Significant genotype × treatment interactions (p ≤ 0.05) were observed for feed intake at week 3, total protein, albumin, cholesterol, triglycerides, HDL, LDL, alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea, creatinine, and glutathione peroxidase (GPx). Feed intake at week 3 showed parallel interaction patterns, with Cobb 500 maintaining higher values than Abor Acres. Total protein was optimized at 1.0-1.5% ginger inclusion in both strains, while albumin responses indicated optimum values at 1.0% in Cobb 500 and 1.5% in Abor Acres. Cholesterol reduction was greatest at 1.0% ginger inclusion in Cobb 500 and 1.5% in Abor Acres. Triglycerides declined markedly at 0.5% inclusion in Abor Acres and 1.0% in Cobb 500. HDL concentrations were lowest at 1.0% inclusion in Cobb 500 and 1.5% inclusion in Abor Acres (19.5 mg/dL), whereas LDL was minimized at 0.5% and 1.5% inclusion levels in Cobb 500 and Abor Acres, respectively. ALT values were lowest at 1.5% inclusion in both strains. Peak ALP, urea, creatinine, and GPx responses occurred at different inclusion levels between strains, indicating genotype-dependent metabolic, hepatic, renal, and antioxidant responses. These findings highlight the importance of genotype-specific dietary ginger supplementation strategies in broiler production.
Matrix metalloproteinase-1 (MMP-1) is a key enzyme that drives extracellular matrix degradation and facilitates breast cancer progression, invasion, and metastasis. This abnormal MMP-1 activity is linked to worse patient outcomes and also improved progression of tumor, which promotes critical therapeutic target. However, recent deep learning approaches are utilized still unable to fully capture molecular structures, cross-domain molecular-protein interactions, and interpretable predictive features. These challenges are addressed by incorporating a novel Contrastive Learning-based Molecular-Protein Deep Kernel Learning (CLM-DKL) in this research to solve constraints during the high-throughput virtual screening of phytochemicals targeting MMP-1. This process is effectively fine-tuned via the Stellar Oscillation Optimizer (SOO). Moreover, the Structure-Enhanced Cross-Interaction Graph Attention Network (SECI-GAT) produces embeddings in a hierarchical manner that capture both intra-molecular structure and molecular-protein interactions. To improve the prediction of molecules with more structurally informative area-focused progress and the alignment of different representations of information, a Multi-View Contrastive Learning (MVCL) with both attentions, such as Structural Entropy Guided Attention (SEGA) and Encoder Attention Fusion (EAF), are utilized. The uncertainty-aware molecular-protein affinity prediction by CLM-DKL, and atom-residue-level contribution scores provided by the proposed Deep Learning Important FeaTures (DeepLIFT) for biological interpretability. Evaluation outcomes achieve ROC-AUC of 0.91, 0.88, 0.90, PR-AUC of 0.87, 0.84, 0.86, for ChEMBL, PubChem BioAssay, and BindingDB, respectively, and F1 scores up to 0.94, demonstrating strong predictive power, stability, and generalization. This integrated framework provides a robust, interpretable, and structurally informed pipeline for discovering potent MMP-1 inhibitors.
Human dietary exposure involves multiple contaminants, raising concerns about "cocktail" effects ignored by single-compound risk assessment. Benzo[a]pyrene (BaP) and aflatoxin B1 (AFB1) are co-occurring food contaminants requiring cytochrome P450-mediated bioactivation. They both form DNA-reactive metabolites and are classified as genotoxic carcinogens. Here, BaP and AFB1 genotoxicity was evaluated individually and in binary combination in metabolically competent HepaRP cells using γH2AX as DNA damage biomarker. Concentrations were selected to reflect physiologically relevant levels (BaP: 0.125-10 µM; AFB1: 0.01-2 µM). Benchmark concentration (BMC) analysis with the PROAST covariate approach allowed deriving potency estimates, predicting mixture effects under dose addition hypothesis. Cells co-exposed to BaP-AFB1 presented a synergistic increase in γH2AX foci number compared to additive model predictions. Mechanistic investigations identified aryl hydrocarbon receptor (AhR) signaling as a key driver of this interaction. First, AhR pharmacological activation with β-naphthoflavone (BNF) recapitulated the effects of BaP, enhancing the genotoxicity induced by AFB1 exposure. In contrast AhR inhibition with CH223191 reduced DNA damage in cells treated with BaP, BaP-AFB1 or BNF-AFB1. Consistent with the metabolic interaction hypothesis, BaP or BNF alone or in mixture with AFB1 increased the expression and activity of CYP1 family enzymes, while CH223191 prevented these responses. Finally, non-genotoxic polycyclic aromatic hydrocarbons (PAHs) as fluoranthene and phenanthrene also enhanced AFB1 genotoxicity. Altogether, these results underline the importance of AhR-dependent metabolic activation in synergistic interaction within mixtures, supporting the necessity of mechanistic knowledge to improve risk assessment, notably for co-exposure to food contaminants.
Gastric cancer (GC) is a very aggressive cancer with a high potential for metastasis. A naturally occurring flavonoid, luteolin (Lut) has metabolic regulation and anti-cancer properties. This work investigates whether Lut modulates glucose metabolism and targets AKR1B1 to prevent the epithelial-mesenchymal transition (EMT) in GC. Lut was used to treat human GC cell lines that were grown in both normal and high-glucose environments. Western blotting and qPCR were used to assess the expression levels of EMT markers and AKR1B1. The CCK-8 assay, the scratch assay, and the Transwell assay were used to measure cell movement, invasion, and proliferation, respectively. Metabolic flux analysis was used to measure fructose and lactate generation in order to assess glycolytic activity. We carried out AKR1B1 overexpression and knockdown experiments to elucidate the functional involvement of AKR1B1 in the action of Lut. In order to fully assess the anticancer effects of Lut and its underlying mechanisms, a high-glucose paradigm in nude mice was developed for in vivo validation. AKR1B1 expression was markedly elevated in GC cells under high-glucose circumstances, and this was accompanied by increased glycolytic activity. At the same time, the cells showed signs of EMT, including enhanced migratory and invasion capacities. Subsequent research showed that Lut therapy substantially alleviated anomalies in glucose metabolism by drastically suppressing AKR1B1 expression, downregulating important glycolytic enzymes, and reducing the generation of fructose and lactate. Additionally, by boosting E-cadherin expression, lowering vimentin and N-cadherin levels, and preventing cell migration and invasion, luteolin dramatically corrected high-glucose-induced EMT. The inhibitory effects of luteolin on glycolysis and EMT were largely offset by AKR1B1 overexpression, according to functional experiments; on the other hand, inhibition of AKR1B1 considerably reduces AKT pathway activation, which suppresses the malignant phenotype of gastric cancer cells. Together, these findings show how important AKR1B1 is to this regulatory axis. According to these results, luteolin inhibits high-glucose-induced metabolic reprogramming and EMT in gastric cancer cells via acting on AKR1B1, which lowers the cells' capacity for metastasis. Lut may reduce the metabolic dysregulation brought on by hyperglycemia by blocking AKR1B1, which in turn prevents GC cells from undergoing the epithelial-mesenchymal transition. This discovery highlights Lut as a potential metabolic therapeutic drug and offers mechanistic insight into the evolution of gastric cancer caused by hyperglycemia.
Thorium is an extremely radioactive and hazardous material and there is increasing concern over the environmental impact of thorium on the health of soils. There have been a limited number of investigations into phytotoxicity, phytoremediation, and the mechanisms of thorium absorption by woody plants. The purpose of this review is to assess the impact of thorium on soil's physical, chemical, and biological characteristics and quantify how much thorium is available for woody plant uptake and how much thorium will be moved around within different parts of the plant. This review also examines the bioavailability, mobility and root uptake of thorium by woody plants. The extent to which woody plants absorb thorium has been reviewed according to three activity concentration rates: low, moderate and high. Furthermore, literature was reviewed and analyzed as to how woody plant absorption may be influenced by having elevated quantities of thorium in the soil, which exceeds the toxic threshold. Thorium adversely affects all living things through hindering seed germination, reducing plant growth, inhibiting photosynthetic rate, and reducing the plants' ability to absorb nutrients; thorium has not been demonstrated to have any biological function. Woody Plants can employ several methods to combat the negative effects of thorium, such as producing phytochelatins, utilizing multiple compartments for storing thorium, and producing many different types of enzymes that function as antioxidants. Therefore, there are numerous technologically based and microbially based methods available for remediating thorium-contaminated soils, one of which is the use of specific plant species as remedial agents. Finally, a thorough review of literature was performed to identify all native woody plants that are best adapted for phytoremediation of thorium-contaminated soils and the mechanisms used by these native woody plant species to protect themselves from the toxic effects of thorium contamination were determined.
Antibody Fc regions have important roles in clearance of Plasmodium falciparum-infected erythrocytes. One such role is engagement with Fcγ receptors on host leukocytes. In the case of FcγRIIIa and b, this interaction is greatly enhanced when the IgG glycan is afucosylated. In this study, we used a fucose-sensitive enzyme-linked immunosorbent assay (FEASI) to assess afucosylation in IgG specific for placental malaria protein VAR2CSA from n = 139 malaria-exposed pregnant Malawian women, correlating these data with mass spectrometry-based analysis. Furthermore, we measured the effect of afucosylation on Fc-mediated leukocyte functions, using both plasma and a monoclonal antibody, PAM2.8, with varying levels of afucosylation. There were significantly higher levels of VAR2CSA-specific IgG afucosylation in women with no placental malaria measured by FEASI (P < .0001), which correlated strongly with mass spectrometry analysis (R = 0.8, P < .0001). In addition, highly afucosylated IgG mediated significantly greater neutrophil phagocytosis of antigen-coated beads and induction of NK cell degranulation by infected erythrocytes. Afucosylated IgG to VAR2CSA, measured by FEASI or mass spectrometry, was a correlate of protection from placental malaria, and afucosylated IgG activated NK cells and neutrophils. Naturally acquired or therapeutic afucosylated IgG antibody could have a role in protection from malaria infection.
The worldwide spread of antimicrobial resistance (AMR) is a considerable challenge to global health, resulting in a substantial depletion of human and material resources. Conventional antimicrobial agents are often ineffective against contemporary resistant strains, making the identification of novel drug targets a priority in antimicrobial development. Inhibition of the bacterial type II fatty acid synthesis (FAS-II) pathway is recognized as an effective antimicrobial strategy. β-Ketoacyl-ACP synthase III (FabH), a crucial enzyme in the FAS-II, presents potential as a target for next-generation antimicrobial agents. In recent years (2006-2025), research on synthetic small-molecule inhibitors targeting FabH has attracted widespread attention among scientists. However, reviews on FabH inhibitors remain scarce, with reports scattered across the literature. This paper outlines the attributes of FabH in various bacterial species and compiles the currently documented synthetic inhibitors. It examines the inhibitory effects of various core structures on FabH and analyzes the structure-activity relationships of specific compounds, including triazoles, carbazoles, indoles, and pyrazoles, etc. The goal is to offer innovative perspectives for the future development and formulation of antibacterial agents targeting FabH.
Hypertension is a risk factor for cognitive impairment, but the mechanistic link between hypertension and cognitive decline remains unclear. We find that captopril failed to protect against synaptic damage in spontaneously hypertensive rats. In contrast, the hazelnut-derived peptide YYLLVR exerted a dual-modulating effect by activating Angiotensin-converting enzyme 2 (ACE2), simultaneously lowering blood pressure and ameliorating cognitive impairment. YYLLVR reduced mean arterial pressure by 43.18 mmHg (60 mg/kg body weight per day for 4 weeks, P < 0.05). In behavioral tests, it reduced escape latency by 11.29 s and alleviated hippocampal neuronal damage (P < 0.05). YYLLVR inhibited ACE, activated ACE2, suppressed neuroinflammation via glial inactivation, reduced pathological p-tau, and enhanced synaptic plasticity. It also ameliorated mitochondrial dysfunction and oxidative stress across multiple cell lines (100 μM). Molecular dynamics confirmed stable binding of YYLLVR to ACE2. This study establishes a nutritional intervention framework for ACE2-activating peptides against hypertension-induced cognitive impairment.
The discovery of biomolecular condensates, driven by liquid-liquid phase separation of intrinsically disordered proteins has significant impacts on both fundamental and applied science and engineering. Although most studies on biomolecular condensates focus on intrinsically disordered structures, research on the role of molecular ordering remains largely unexplored, however is beneficial for gaining new mechanistic understanding and further expand the design space of peptides for constructing functional condensates. Toward this goal, we conducted systematic studies on how molecular ordering impacts the phase behaviors of peptides using multidomain peptides (MDPs) as a model system. MDPs were designed using a molecular frustration principle in which parts of the peptides favored β-sheet assembly and parts favored disassembly. Through programming of each domain, it is evident that the phase behavior of MDPs is largely dictated by the secondary structure, and partially folded β-sheet plays a key role in driving MDPs to form condensates. We also discovered complex coacervates formed by MDPs and synthetic anionic polymers, which exhibited dramatically improved stability. Furthermore, we show enzyme-triggered condensation can be achieved using phosphorylated MDPs as the molecular precursor and alkaline phosphatase as a molecular switch, highlighting the potential of these materials for bacterial imaging and antimicrobial therapy development.