Main Binding Site Research Articles

Effect of conditioners used for sludge dewatering on the adsorption performance of sludge-derived adsorbent

Converting waste activated sludge into adsorbent is a promising alternative for sludge valorization. While various conditioners are widely employed in sludge conditioning and dewatering processes, the influence of these conditioners on the properties of sludge derived adsorbents remains largely unexplored. Four conditioners, including FeCl3, CaO, polyacrylamide, FeSO4·7 H2O/Na2S2O8, were used for sludge conditioning, and the sludge dewaterability and the adsorption performance of the resultant adsorbents were evaluated. The sludge dewaterability was markedly improved by the conditioners in a descending order as polyacrylamide, FeCl3, FeSO4·7 H2O/Na2S2O8, and CaO. The type of conditioner highly influenced the adsorption ability of the sludge adsorbent: FeSO4·7 H2O/Na2S2O8 enhanced the adsorption ability, while others deteriorated it. Infrared spectra, pore size distribution, and micromorphology analysis of the adsorbents suggest that: FeCl3 reduced mesopores through strong electrostatic interactions; CaO acted as a physical barrier for adsorption; polyacrylamide reduced the adsorbent surface area through hydrogen bond-driven bridging and potentially forming polymeric barrier; FeSO4·7 H2O/Na2S2O8 disrupted the sludge flocs and exposed abundant adsorption sites, promoting the adsorbent performance markedly. FeSO4·7 H2O/Na2S2O8 conditioning was thus beneficial to both sludge dewatering and adsorption. Adsorption of methylene blue by FeSO4·7 H2O/Na2S2O8-conditioned sludge adsorbent was best described by the pseudo-second-order model and Langmuir model, indicating the dominance of monolayer chemisorption. The adsorption mainly occurred in the mesopores of the adsorbent, with hydroxyl group and carbonyl in carboxyl group as the main binding sites. Both film and pore diffusion contributed to the adsorption rate, with pore diffusion serving as the main rate limiting step. Results highlight the beneficial effect of destructive conditioning method on sludge adsorbent performance.

Functional characterization of large yellow croaker (Larimichthys crocea) Peroxiredoxin IV (PrxIV) gene promoter

Peroxiredoxin IV (PrxIV), which possesses an N-terminal signal peptide, is the only secretable protein in Prx family. PrxIV can protect cells against reactive oxygen species (ROS) and act as a DAMP to promote infection-independent immune response. However, the characterization and regulation of promoters of PrxIV genes are rarely reported. In this study, a 1511-bp 5′-flanking sequence of large yellow croaker (Larimichthys crocea) PrxIV (LcPrxIV) was cloned and characterized. DNA truncation combined with luciferase activity assay revealed that a fragment of −781/+20 contained in the plasmid LcPrxIV-P3 exhibited the highest promoter activity. It could initiate the luciferase expression up to 44.6-fold when compared to control plasmid pGL3-Basic. TFSEARCH analysis revealed many recognizing sequences of transcriptional factors exist within this 1511-bp sequence, including Foxo and CREB. Altogether, four putative binding sites located in three recognizing sequences of CREB were identified. Notably, co-transfection of LcPrxIV-P3 with LcCREB led to a significant 2.48-fold increase of the LcPrxIV-P3 promoter activity (P<0.01). Furthermore, the mutation at putative binding sites A, B, and all four sites of CREB in the LcPrxIV-P3 caused the significant decrease of activation on LcPrxIV-P3 promoter activity, suggesting these two sites may be the main binding sites of CREB in LcPrxIV promoter. In addition, the oxidative stress caused by hydrogen peroxide, rather than immune stimuli such as Poly (I: C), LPS, LTA, or PGN could lead to the elevation of LcPrxIV-P3 promoter activity. When the concentration of hydrogen peroxide reached 500 μM, the promoter activity of LcPrxIV-P3 could be up-regulated to 1.47-fold, which was extremely significantly different from the control (P<0.001). These results help to elucidate the regulatory mechanisms of LcPrxIV gene expression, and the role of LcPrxIV in protecting cells against oxidative stress or in oxidoreduction-dependent signal transduction.

CheB localizes to polar receptor arrays during repellent adaptation.

Adaptation of the response to stimuli is a fundamental process for all organisms. Here, we show that the adaptation enzyme CheB methylesterase of Escherichia coli assembles to the ON state receptor array after exposure to the repellent l-isoleucine and dissociates from the array after adaptation is complete. The duration of increased CheB localization and the time of highly clockwise-biased flagellar rotation were similar and depended on the strength of the stimulus. The increase in CheB at the receptor array and the decrease in cytoplasmic CheB were both ~100 molecules, which represents 15 to 20% of the total cellular content of CheB. We confirmed that the main binding site for CheB in the ON state array is the P2 domain of phosphorylated CheA, with a second minor site being the carboxyl-terminal pentapeptide of the serine chemoreceptor. Thus, we have been able to quantify the regulation of the signal output of the receptor array by the intracellular dynamics of an adaptation enzyme.

Atomistic Insights into the Stabilization of TDP-43 Protofibrils by ATP.

The aberrant accumulation of the transactive response deoxyribonucleic acid (DNA)-binding protein of 43 kDa (TDP-43) aggregates in the cytoplasm of motor neurons is the main pathological hallmark of amyotrophic lateral sclerosis (ALS). Previous experiments reported that adenosine triphosphate (ATP), the universal energy currency for all living cells, could induce aggregation and enhance the folding of TDP-43 fibrillar aggregates. However, the significance of ATP on TDP-43 fibrillation and the mechanism behind it remain elusive. In this work, we conducted multiple atomistic molecular dynamics (MD) simulations totaling 20 μs to search the critical nucleus size of TDP-43282-360 and investigate the impact of ATP molecules on preformed protofibrils. The results reveal that the trimer is the critical nucleus for TDP-43282-360 fibril formation and the tetramer is the minimal stable nucleus. When ATP molecules bind to the TDP-43282-360 trimer and tetramer, they can consolidate the TDP-43282-360 protofibrils by increasing the content of the β-sheet structure and promoting the formation of hydrogen bonds (H-bonds). Binding site analyses show that the N-terminus of TDP-43282-360 protofibrils is the main binding site of ATP, and R293 dominates the direct binding of ATP. Further analyses reveal that the π-π, cation-π, salt bridge, and H-bonding interactions together contribute to the binding of ATP to TDP-43282-360 protofibrils. This study decoded the detailed stabilization mechanism of protofibrillar TDP-43282-360 oligomers by ATP, and may provide new avenues for the development of drug design against ALS.

A novel strategy for sequential separation of Th(IV) from highly acidic wastewater based on solid phase extraction

The potential applications and advantages of thorium(Th) in the development of the nuclear industry make the effective separation of Th(IV) crucial for depleted fuel management and environmental safety. However, selectively capturing of Th(IV) from high-concentration nitric acid remains a challenging task. In this work, a novel solid-phase extraction resin (TOPO/XAD-7) was successfully synthesized by vacuum impregnation of trioctylphosphine oxide (TOPO) into an acrylic-based polymer (XAD-7 resin). It exhibited excellent adsorption abilities for Th(IV), including wide highly concentration acid tolerance (0.01–6 mol L−1 HNO3), high adsorption capacity of 59.79 mg g−1, fast adsorption kinetic (< 60 min) and super elevated selectivity. The adsorption mechanism of the resin involves neutral chelation extraction, with the PO group being the main adsorption binding site. Additionally, the resin was demonstrated good reusability and stability under extreme conditions (high acidity, high temperature and high radiation). Furthermore, dynamic column experiments conducted using an automated separation system validated the practical application of TOPO/XAD-7 resin for the separation of Th(IV) and U(VI)/Th(IV) in wastewater, achieving high element recovery (> 99.5%) and a high U(VI) decontamination factor (3.5 × 105). This work confirms the excellent potential of TOPO/XAD-7 resin for the separation and enrichment of Th(IV) in nuclear wastewater.

Association mechanism of bicalutamide and human serum albumin for potential clinical implications.

The binding mechanism of molecular interaction between bicalutamide and human serum albumin (HSA) in a pH7.4 phosphate buffer was studied using various spectroscopic techniques in combination with molecular modeling. Fluorescence data revealed that the fluorescence quenching of HSA by bicalutamide was a static quenching procedure. The binding constants and number of binding sites were evaluated at different temperatures. The thermodynamic parameters, ΔH and ΔS, were calculated to be 4.30 × 104J·mol-1 and 245 J·mol-1·K-1, respectively, suggesting that the binding of bicalutamide to HSA was driven mainly by hydrophobic interactions and hydrogen bonds. The displacement studies indicated neither Sudlow's site I nor II but subdomain IB as the main binding site for bicalutamide on HSA. The binding distance between bicalutamide and HSA was determined to be 3.54 nm based on the Förster theory. Analysis of circular dichroism, synchronous, and 3D fluorescence spectra demonstrated that HSA conformation was slightly altered in the presence of bicalutamide.

Metadynamics Study of Lipid-Mediated Antibacterial Toxin Binding to the EmrE Multiefflux Protein.

EmrE is a bacterial efflux protein in the small multidrug-resistant (SMR) family present in Escherichia coli. Due to its small size, 110 residues in each dimer subunit, it is an ideal model system to study ligand-protein-membrane interactions. Here in our work, we have calculated the free energy landscape of benzyltrimetylammonium (BTMA) and tetraphenyl phosphonium (TPP) binding to EmrE using the enhanced sampling method-multiple walker metadynamics. We estimate that the free energy of BTMA binding to EmrE is -21.2 ± 3.3 kJ/mol and for TPP is -43.6 ± 3.8 kJ/mol. BTMA passes through two metastable states to reach the binding pocket, while TPP has a more complex binding landscape with four metastable states and one main binding site. Our simulations show that the ligands interact with the membrane lipids at a distance 1 nm away from the binding site which forms a broad local minimum, consistent for both BTMA and TPP. This site can be an alternate entry point for ligands to partition from the membrane into the protein, especially for bulky and/or branched ligands. We also observed the membrane lipid and C-terminal 110HisA form salt-bridge interactions with the helix-1 residue 22LysB. Our free energy estimates and clusters are in close agreement with experimental data and give us an atomistic view of the ligand-protein-lipid interactions. Understanding the binding pathway of these ligands can guide us in future design of ligands that can alter or halt the function of EmrE.

Dissecting molecular mechanisms underlying the inhibition of β-glucuronidase by alkaloids from Hibiscus trionum: Integrating in vitro and in silico perspectives

β-Glucuronidase, a crucial enzyme in drug metabolism and detoxification, represents a promising target for therapeutic intervention due to its potential to modulate drug pharmacokinetics and enhance therapeutic efficacy. Herein, we assessed the inhibitory potential of phytochemicals from Hibiscus trionum against β-glucuronidase. Grossamide and grossamide K emerged as the most potent β-glucuronidase inhibitors with IC50 values of 0.73 ± 0.03 and 1.24 ± 0.03 μM, respectively. The investigated alkaloids effectively inhibited β-glucuronidase-catalyzed PNPG hydrolysis through a noncompetitive inhibition mode, whereas steppogenin displayed a mixed inhibition mechanism. Molecular docking analyses highlighted grossamide and grossamide K as inhibitors with the lowest binding free energy, all compounds successfully docked into the same main binding site occupied by the reference drug Epigallocatechin gallate (EGCG). We explored the interaction dynamics of isolated compounds with β-glucuronidase through a 200 ns molecular dynamics (MD) simulation. Analysis of various MD parameters revealed that grossamide and grossamide K maintained stable trajectories and demonstrated significant energy stabilization upon binding to β-glucuronidase. Additionally, these compounds exhibited the lowest average interaction energies with the target enzyme. The MM/PBSA calculations further supported these findings, showing the lowest binding free energies for grossamide and grossamide K. These computational results are consistent with experimental data, suggesting that grossamide and grossamide K could be potent inhibitors of β-glucuronidase.

Surface properties of activated carbon fibers obtained from polyacrylonitrile and methyl acrylate: Experimental and simulation studies for lead and acid blue 25 dye adsorption from water

Activated carbon fibers were prepared using a methyl acrylate/polyacrylonitrile (MA/PAN) system and H3PO4 activation. These fibers were assessed to adsorb AB25 and Pb2+ from aqueous solutions, and DFT calculations were carried out to determine the main surface interactions that caused the removal of both pollutants. The results showed that AB25 and Pb2+ adsorption on these activated carbon fibers was endothermic, with maximum experimental adsorption capacities of 0.012 − 0.023 and 0.248 − 0.306 mmol/g at 20 – 40 °C. Pb2+ adsorption properties of these fibers outperformed those of other carbonaceous materials utilized in the depollution of metallic cations, whereas their removal effectiveness was less competitive to separate AB25 dye molecules. A double-site monolayer model was applied to correlate the experimental Pb2+ isotherms, while AB25 adsorption on activated carbon fibers was simulated using a one-site multilayer model. Oxygenated acidic (i.e., carboxylic and phenolic) and phosphate-carbon functional groups were the main binding sites to adsorb these pollutants. The adsorption energies of the tested systems, the identification of the most feasible binding positions of the tested adsorbates on the activated carbon fiber surface, and the role of hydrogen bonding in the corresponding adsorption mechanisms were discussed using DFT calculations. These results contribute to the valorization and application of engineered materials such as activated carbon fibers for water depollution.

Deciphering the interplay between wastewater compositions and oxytetracycline in recovered struvite: Unveiling mechanisms and introducing control strategies

Struvite recovery from wastewater offers a sustainable phosphorus and nitrogen source, yet it harbors the challenge of variable antibiotic residues, notably oxytetracycline (OTC), increasing the ecological risk during subsequent use. Despite the need, mechanisms behind these residues and regulatory solutions remain obscure. We characterized OTC in recovered struvite and showed that increased dissolved organic matter (DOM) enhanced OTC accumulation, while PO43- suppressed it. NH4+ modulated OTC levels through the saturation index (SI), with a rise in SI significantly reducing OTC content. Additionally, excess Mg2+ formed complexes with OTC and DOM (humic acid, HA), leading to increased residue levels. Complexation was stronger at higher pH, whereas electrostatic interactions dominated at lower pH. The primary binding sites for antibiotics and DOM were Mg-OH and P-OH groups in struvite. OTC's dimethylamino, amide, and phenolic diketone groups primarily bound to struvite and DOM, with the carboxyl group of DOM serving as the main binding site. Mg2+ complexation was the primary pathway for OTC transportation, whereas electrostatic attraction of PO43- dominated during growth. Controlling magnesium (Mg) dosage and adjusting pH were effective for reducing OTC in recovered products. Our findings provided insights into the intricate interactions between struvite and antibiotics, laying the groundwork for further minimizing antibiotic residues in recovered phosphorus products.

Experimental and DFT calculation study on the efficient removal of high fluoride wastewater from metallurgical wastewater by kaolinite

Fluoride pollution and water scarcity are urgent issues. Reducing fluoride concentration in water is crucial. Kaolinite has been used to study adsorption and fluoride removal in water and to characterize material properties. The experimental results showed that the adsorption capacity of kaolinite decreased with increasing pH. The highest adsorption of fluoride occurred at pH 2, with a capacity of 11.1 mg/g. The fluoride removal efficiency remained high after four regeneration cycles. The fitting results with the Freundlich isotherm model and the external diffusion model showed that the non-homogeneous adsorption of kaolinite fit the adsorption behavior better. Finally, the adsorption mechanism was analyzed by FT-IR and XPS. The binding energies of various adsorption sites and the chemical adsorption properties of atomic states were discussed in relation to DFT calculations. The results showed that Al and H sites were the main binding sites, and the bonding stability for different forms of fluoride varies, with the size of Al–F (−7.498 eV) > H–F (−6.04 eV) > H–HF (−3.439 eV) > Al–HF (−3.283 eV). Furthermore, the density of states and Mulliken charge distribution revealed that the 2p orbital of F was found to be active in the adsorption process and was the main orbital for charge transfer.

#2765 Medium chain fatty acids are potent binding competitors to improve protein-bound uremic toxin clearance in hemodialysis

Abstract Background and Aims Protein-bound uremic toxins (PBUTs) remain a concerning burden in patients with end-stage renal disease (ESRD) since their removal during hemodialysis is limited due to their tight binding to albumin. By increasing their circulating free fraction during HD, “binding competitors” of albumin could be used to increase their dialytic clearance. Medium chain fatty acids (MCFAs, 6 to 12 carbon length) are potent candidates for that purpose. The present study aims to investigate whether MCFAs could serve as potential binding competitors of albumin to enhance the removal of PBUTs, specifically IS and p-CS, during hemodialysis. Method Hexanoate (C6), octanoate (C8), and decanoate (C10) were purchased from Sigma-Aldrich. The binding of MCFAs to albumin was investigated in silico and using fluorescent probes: warfarin (specific of Sudlow's site I) and dansylsarcosine (specific of Sudlow's site II). Indoxyl-sulfate (IS, final concentration 212 µM) and p-cresyl sulfate (p-CS, final concentration 250 µM) were added to a 600 µM bovine serum albumin-phosphate buffered saline solution. The free fractions of IS and p-CS were assayed with or without MCFAs (1-2 mM) using ultrafiltration devices in this solutions and in patients uremic plasma. To mimic the removal of PBUTs during a HD session, batches of 2 liters of fresh bovine blood were loaded with IS and p-CS (final concentration 200 µM). A 2-hours closed-loop HD session was performed using a Fresenius 5008 CorDiax HD generator (Fresenius, Germany). A solution of 224 mM of octanoate was perfused at the rate of 150 µL/min; A solution of saline 0.9% was used as a control. One milliliter of blood was sampled every 15 minutes through the arterial sampling port and concentration of IS and p-CS were assayed by HPLC coupled with fluorescence detection. The hemolytic effect of MCFAs was evaluated in vitro by assaying the free hemoglobin concentration. Results According to the in silico prediction, the binding was predicted to be almost complete for MCFAs ≥ C8 (Fig. 1). The enhanced protein binding appeared to be inversely correlated with water solubility, which became particularly low for MCFAs ≥ C10. Octanoate and decanoate were the more prone to displace dansylsarcosine from Sudlow's site II of albumin (which is the main binding site of IS and p-CS). In vitro, the incubation with 2 mM of octanoate and decanoate increased the free fraction of PBUTs from 12% to 53% for p-CS (4.4 fold, p &amp;lt; 0.05) and from 11% to 45% for IS (4.1 fold, p &amp;lt; 0.05). Octanoate was also able to increase the free fraction of PBUTs in patients uremic plasma (1.6-fold for IS, p &amp;lt; 0.05 and 4.8 fold for p-CS, &amp;lt;0.01). The per-dialytic infusion of octanoate significantly increased the fractional removal of p-CS (from 38% to 88%, p &amp;lt; 0.001) and IS (from 36% to 91%, p &amp;lt; 0.001). No significant hemolysis was observed for concentration of MCFAs lower than 2 mmol/L. Conclusion MCFAs and especially octanoate and decanoate are serious candidates to displace the binding of PBUTs such as p-CS and IS. The per-dialytic administration of octanoate significantly increased the removal of PBUTs and could constitute a new strategy to prevent their accumulation in end-stage kidney disease patients. Due to their good safety profile, MCFAs could be better tolerated than other chemical compounds that has already been tested in a clinical setting such as ibuprofen. Further in vivo studies are however needed to carefully evaluate this potentially new therapeutic option.

Synthesis and Reactions of 3‐Amino‐5,6‐Dimethyl‐l,2,4‐Triazine: DFT Studies, ADME Assay, and Molecular Docking on the new l,2,4‐Triazine Derivatives as Anticancer Agents.

AbstractTwo effective one‐pot synthesis procedures were proposed for the synthesis of 3‐amino‐5,6‐dimethyl‐l,2,4‐triazine under free solvent conditions; these methods included fusing the monohydrazide of biacetyl with either methyl carbamimidothioate (Method B) or cyanamide (Method C). Simple reaction conditions, an easy work‐up process, and facile separation characterize the ecologically friendly fusion process used in techniques B and C. The results from methods B and C indicated the existence of amino‐imino prototropic tautomerism. The triazine in method C had a different melting point and had unusual mass spectral behavior, suggesting that it was in the imino‐tautomer form, as opposed to the amino‐tautomer form discovered in procedure B. The strong electron‐withdrawing activity of the triazine ring appears to deactivate the 3‐amino group, which in turn triggers the production of additional triazine derivatives (3–12), with yields ranging from 54% to 82%. To characterize the structure of the newly synthesized compounds, 1H, 13C‐NMR, and mass spectrometry were employed. The compounds that were created underwent testing at the National Cancer Institute. According to the screening results, conjugates 9 and 10 were successful against most of the cancer cell line subpanels. With the use of DFT, we conducted theoretical studies on compounds 9 and 10. Topological studies, including ELF, LOL, and RDG, were used to identify the main binding sites and weak interactions of the target derivatives. The synthesized compounds that have promising properties are subjected to a pharmacological examination using Swiss‐ADME. Cox‐1 (PDB ID: 3KK6) is the target of molecular docking experiments that aim to identify the anticancer active site of the derivatives of interest.

Molecular insights into linkages among free-floating macrophyte-derived organic matter, the fate of antibiotic residues, and antibiotic resistance genes

Macrophyte rhizospheric dissolved organic matter (ROM) served as widespread abiotic components in aquatic ecosystems, and its effects on antibiotic residues and antibiotic resistance genes (ARGs) could not be ignored. However, specific influencing mechanisms for ROM on the fate of antibiotic residues and expression of ARGs still remained unclear. Herein, laboratory hydroponic experiments for water lettuce (Pistia stratiotes) were carried out to explore mutual interactions among ROM, sulfamethoxazole (SMX), bacterial community, and ARGs expression. Results showed ROM directly affect SMX concentrations through the binding process, while CO and N-H groups were main binding sites for ROM. Dynamic changes of ROM molecular composition diversified the DOM pool due to microbe-mediated oxidoreduction, with enrichment of heteroatoms (N, S, P) and decreased aromaticity. Microbial community analysis showed SMX pressure significantly stimulated the succession of bacterial structure in both bulk water and rhizospheric biofilms. Furthermore, network analysis further confirmed ROM bio-labile compositions as energy sources and electron shuttles directly influenced microbial structure, thereby facilitating proliferation of antibiotic resistant bacteria (Methylotenera, Sphingobium, Az spirillum) and ARGs (sul1, sul2, intl1). This investigation will provide scientific supports for the control of antibiotic residues and corresponding ARGs in aquatic ecosystems.

Typical endocrine disruptors diethylstilbestrol and its analogues non-covalently bind to human serum albumin

Endocrine disruptors have received widespread attention from the scientific community. However, the structure-activity relationship of the binding between endocrine disruptors and proteins is still unclear. Therefore, this study investigated the structure-activity relationship between human serum albumin (HSA) and three types of estrogens, hexestrol (HX), diethylstilbestrol (DES), and dienestrol (DIS), which have slightly different structures. The binding constants of HX, DES and DIS with HSA at 298 K were 1.40×104, 3.84×104 and 4.46×104 M−1, respectively. The data of thermodynamic parameters confirmed that the entropy and enthalpy of the estrogens with HSA were positive, indicating that the binding was dominated by hydrophobic interaction. The results from photochemistry and electrochemistry showed that the binding of estrogens to HSA was more tightly regulated by the conjugation effect. The conjugation effect not only changed the binding behavior of estrogen to HSA, but also the selectivity of the binding sites of the three estrogens. These changes caused the main binding sites to become different, while also affecting the local conformation of HSA. This article provides theoretical guidance for predicting the biological safety of analogues of endocrine disruptors at the protein level.

Exploring the impact of malondialdehyde and 4-hydroxy-2-nonena on myofibrillar protein structure: A proteomic analysis

In this investigation, the impacts of malondialdehyde (MDA), 4-hydroxy-2-nonena (HNE), and their combinations on the structural characteristics of myofibrillar proteins (MP) were examined using endogenous fluorescence, Fourier transform infrared spectrum (FTIR) and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). The potential binding mechanisms were also explored using LC-MS/MS and proteomics techniques. The findings indicated that various reactive carbonyl compounds (RCCs) displayed distinct oxidation performances in the oxidation of myofibrillar proteins. Samples containing HNE (HMP and MHMP groups) exhibited a more pronounced ability to oxidize MP than those containing MDA, as evidenced by their higher carbonyl content and lower sulfhydryl content. The MHMP group demonstrated the greatest surface hydrophobicity and Schiff base content but the lowest fluorescence intensity, suggesting that MDA and HNE synergistically altered MP conformation. Moreover, the alteration by RCCs notably modified the secondary structure of MP. Among these, HMP triggered the unfolding of MP molecules, resulting in the lowest α-helix and highest β-sheet ratio. The SDS‒PAGE results suggested that MDA oxidation promoted the crosslinking of MP molecules. At the same time, HNE caused the degradation of MP, as indicated by the decrease in the intensity of the high-molecular-weight bands. Proteomic analysis suggested that lysine residues distributed in the tail region of myosin were the main binding sites for MDA, whereas HNE preferentially modified methionine located at the myosin head. Additionally, HNE-induced protein degradation dominated RCC-induced oxidation reactions. These findings provide deeper insights into the mechanisms underlying quality changes induced by lipid oxidation products in MP-based food systems.

Functional dissection of the spike glycoprotein S1 subunit and identification of cellular cofactors for regulation of swine acute diarrhea syndrome coronavirus entry.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel porcine enteric coronavirus, and the broad interspecies infection of SADS-CoV poses a potential threat to human health. This study provides experimental evidence to dissect the roles of distinct domains within the SADS-CoV spike S1 subunit in cellular entry. Specifically, we expressed the S1 and its subdomains, S1A and S1B. Cell binding and invasion inhibition assays revealed a preference for the S1B subdomain in binding to the receptors on the cell surface, and this unknown receptor is not utilized by the porcine epidemic diarrhea virus. Nanoparticle display demonstrated hemagglutination of erythrocytes from pigs, humans, and mice, linking the S1A subdomain to the binding of sialic acid (Sia) involved in virus attachment. We successfully rescued GFP-labeled SADS-CoV (rSADS-GFP) from a recombinant cDNA clone to track viral infection. Antisera raised against S1, S1A, or S1B contained highly potent neutralizing antibodies, with anti-S1B showing better efficiency in neutralizing rSADS-GFP infection compared to anti-S1A. Furthermore, depletion of heparan sulfate (HS) by heparinase treatment or pre-incubation of rSADS-GFP with HS or constituent monosaccharides could inhibit SADS-CoV entry. Finally, we demonstrated that active furin cleavage of S glycoprotein and the presence of type II transmembrane serine protease (TMPRSS2) are essential for SADS-CoV infection. These combined observations suggest that the wide cell tropism of SADS-CoV may be related to the distribution of Sia or HS on the cell surface, whereas the S1B contains the main protein receptor binding site. Specific host proteases also play important roles in facilitating SADS-CoV entry.IMPORTANCESwine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel pathogen infecting piglet, and its unique genetic evolution characteristics and broad species tropism suggest the potential for cross-species transmission. The virus enters cells through its spike (S) glycoprotein. In this study, we identify the receptor binding domain on the C-terminal part of the S1 subunit (S1B) of SADS-CoV, whereas the sugar-binding domain located at the S1 N-terminal part of S1 (S1A). Sialic acid, heparan sulfate, and specific host proteases play essential roles in viral attachment and entry. The dissection of SADS-CoV S1 subunit's functional domains and identification of cellular entry cofactors will help to explore the receptors used by SADS-CoV, which may contribute to exploring the mechanisms behind cross-species transmission and host tropism.

Metal-Organic Frameworks Possessing Suitable Pores for Xe/Kr Separation.

Adsorption separation of the Xe/Kr mixture remains a tough issue since Xe and Kr have an inert nature and similar sizes. Here we present a chlorinated metal-organic framework (MOF) [JXNU-19(Cl)] and its nonchlorinated analogue (JXNU-19) for Xe/Kr separation. The two isostructural MOFs constructed from the heptanuclear cobalt-hydroxyl clusters bridged by organic ligands are three-dimensional structures. Detailed contrast of the Xe/Kr adsorption separation properties of the MOF shows that significantly enhanced Xe uptakes and Xe/Kr adsorption selectivity (17.1) are observed for JXNU-19 as compared to JXNU-19(Cl). The main binding sites for Xe in the MOF revealed by computational simulations are far away from the chlorine sites, suggesting that the introduction of the chlorine groups results in the unfavorable Xe adsorption for JXNU-19(Cl). The optimal pores, high surface area, and multiple strong Xe-framework interactions facilitate the effective Xe/Kr separation for JXNU-19.

Effect of pyrolysis temperature on the binding characteristics of DOM derived from livestock manure biochar with Cu(II).

Biochar-derived dissolved organic matter (BDOM) has the potential to influence the environmental application of biochar and the behavior of heavy metals. In this study, the binding properties of BDOM derived from livestock manure biochar at different pyrolysis temperatures with Cu(II) were investigated based on a multi-analytical approach. The results showed that the DOC concentration, aromatics, and humification degree of BDOM were higher in the process of low pyrolysis of biochar. The pyrolysis temperature changed the composition of BDOM functional groups, which affected the binding mechanism of BDOM-Cu(II). Briefly, humic-like and protein-like substances dominated BDOM-Cu(II) binding at low and high pyrolysis temperatures, respectively. The higher binding capacity for Cu(II) was exhibited by BDOM derived from the lower pyrolysis temperature, due to the carboxyl as the main binding site in humic acid had high content and binding ability at low-temperature. The amide in proteins only participated in the BDOM-Cu(II) binding at high pyrolysis temperature, and polysaccharides also played an important role in the binding process. Moreover, the biochar underwent the secondary reaction at certain high temperatures, which led to condensation reaction of the aromatic structure and the conversion of large molecules into small molecules, affecting the BDOM-Cu(II) binding sites.

Increasing the polarity of β-lapachone does not affect its binding capacity with bovine plasma protein

Despite ortho-quinones showing several biological and pharmacological activities, there is still a lack of biophysical characterization of their interaction with albumin - the main carrier of different endogenous and exogenous compounds in the bloodstream. Thus, the interactive profile between bovine serum albumin (BSA) with β-lapachone (1) and its corresponding synthetic 3-sulfonic acid (2, under physiological pH in the sulphonate form) was performed. There is one main binding site of albumin for both β-lapachones (n ≈ 1) and a static fluorescence quenching mechanism was proposed. The Stern-Volmer constant (KSV) values are 104 M−1, indicating a moderate binding affinity. The enthalpy (−3.41 ± 0.45 and − 8.47 ± 0.37 kJ mol−1, for BSA:1 and BSA:2, respectively) and the corresponding entropy (0.0707 ± 0.0015 and 0.0542 ± 0.0012 kJ mol−1 K−1) values indicate an enthalpically and entropically binding driven. Hydrophobic interactions and hydrogen bonding are the main binding forces. The differences in the polarity of 1 and 2 did not change significantly the affinity to albumin. In addition, the 1,2-naphthoquinones showed a similar binding trend compared with 1,4-naphthoquinones.