Reduction Of Disulfide Bonds Research Articles

PAK1 Is Involved in the Spindle Assembly during the First Meiotic Division in Porcine Oocytes.

P21-activated kinase 1 (PAK1), as a member of the PAK family, has been implicated in various functions during somatic mitosis; however, less is known about its role during oocyte meiosis. Herein, we highlight the indispensable role of PAK1 in regulating spindle assembly and cell cycle progression during the first meiotic division of porcine oocytes. First, we found that the activated PAK1 expressed dynamically, and its subcellular localization was tightly associated with the spindle dynamics during meiosis in porcine oocytes. Specific inhibition of PAK1 activity by inhibitor targeting PAK1 activation-3 (IPA-3) led to impaired extrusion of the first polar body (PB1); with most of the IPA-3-treated oocytes arrested at germinal vesicle breakdown (GVBD) and subjected to failure of bipolar spindle formation. However, the adverse effects caused by IPA-3 on oocytes could be restored by reducing disulfide bonds between PAK1 and IPA-3 with dithiothreitol (DTT) treatment. Furthermore, the co-immunoprecipitation assay revealed that PAK1 interacted directly with Aurora A and transforming acidic coiled coil 3 (TACC3), providing an additional explanation for the similar localization of Aurora A and activated PAK1. Additionally, inhibiting the activity of PAK1 decreased the expression of p-Aurora A and p-TACC3; however, the reduced activity of Aurora A and TACC3 could be restored by DTT. In conclusion, PAK1 plays a crucial role in the proper assembly of the spindle during the first meiotic division of porcine oocytes, and the regulation of PAK1 is associated with its effects on p-Aurora A and p-TACC3 expression.

OsPDIL1-1 controls ROS generation by modulating NADPH oxidase in developing anthers to alter the susceptibility of floret fertility to heat for rice

High temperature (HT) at meiosis induces heat injury to pollen viability and floret fertility, which is closely associated with HT-induced endoplasmic reticulum (ER) stress and ROS damage in developing anthers. Disulfide isomerase like proteins (PDILs) play an essential role in the formation, reduction, and isomerization of disulfide bonds in nascent secretory proteins for the maintenance of cell viability and ER homeostasis. However, the underlying mechanism by which HT induces ROS burst in rice anthers and its relation to ER stress for the varying existence of PDILs is largely unknown. In this paper, we investigated the action of PDILs in the regulation of heat injury to floret fertility and its association with HT-induced ROS generation in developing anthers under well-controlled climatic conditions. Results showed that knock-down of OsPDIL1-1 by RNAi enhanced the activity of NADPH oxidase and caused the excessive ROS accumulation in developing anthers, consequently the up-grading sensitivity of pollen viability and floret fertility to heat stress. RBOHb is the primary site where HT exposure affected NADPH oxidase activity and triggered ROS generation in rice anthers because OsPDIL1-1 was found to interact with RBOHb in the ER-PM junction. Furthermore, HT exposure triggered the RBOHb-mediated ROS generation in a Ca 2+ -dependent manner, while the induction of HT exposure to ER stress was not necessarily associated with ROS generation derived from NADPH oxidase. • PDIL1-1 was essential for maintaining floret fertility at elevated temperatures. • PDIL1-1 directly interacts with RBOHb to modulate ROS generation in rice anthers. • HT triggered the RBOHb-mediated ROS generation in a Ca 2+ -dependent manner. • HT-induced ER stress was not associated with NADPH oxidase-derived ROS. • PDIL1-1 was found to interact with RBOHb in the ER-PM junction.

Reducing disulfide bonds as a robust strategy to facilitate the self-assembly of cod protein fabricating potential active ingredients-nanocarrier

In this study, a novel method was developed to encapsulate hydrophobic compounds by self-assembly of cod protein (CP) triggered by breaking disulfide bonds. Curcumin (Cur), a representative lipid-soluble polyphenol, was selected as a model to evaluate the potential of CP nanoparticles as novel and accessible nanocarriers. Results showed that the protein structure gradually unfolded with increasing dithiothreitol (DTT) concentration, indicating that S-S cleavage was conducive to forming a looser structure. The resultant unfolded CP exposed more hydrophobic sites, facilitating its interaction with hydrophobic compounds. The encapsulation efficiency (EE) of formed CP-Cur nanoparticles was relatively high, reaching 99.09%, 98.8%, and 89.77% when the mass ratios of CP to Cur were 20:1, 10:1, and 5:1 (w/v), respectively. The hydrophobic interaction, weak van der Waals, and hydrogen bond were the forces contributing to the formation of CP-Cur nanoparticles, whereas the hydrophobic interaction played a crucial role. The CP-Cur complex exhibited increased stability and a homogeneous-stable structural phase. Thus, this research not only proposed a novel and simple encapsulation method of hydrophobic bioactive compounds but also provided a theoretical reference for the application of reductants in food or pharmacy system.

Self-assembling Bioadhesive Inspired by the Fourth Repetitive Sequence of Balanus albicostatus Cement Protein 20kDa (Balcp-20k).

Barnacle cement proteins are multi-protein complexes composed of a series of functionally related synergistic proteins that enable barnacles to adhere strongly and consistently to various underwater substrates. There is no post-translational modification of barnacle cement proteins, which provides a possibility for the synthesis of similar adhesive materials. Balcp-20k has four repetitive sequences with multiple conserved cysteine groups. Whether these repeats are separate functional units and the role of cysteine in adhesion is not clear. In order to investigate the adhesion properties of Balcp-20k, we amplified and expressed R4 (DHLACNAKHPCWHKHCDCFC)4, which is a quadruple repeat of Balcp-20k's fourth repetitive sequence, and S0R4 (DHLASNAKHPSWHKHSDSFS)4, all cysteine of R4 replaced by serine. Analysis showed that R4 had a similar structure to Balcp-20k, and the amyloid fibrils structure formed by self-assembly of R4 played an important role in improving the adhesion strength. The absence of disulfide bonds in S0R4 prevents self-assembly, and the failure of self-assembly after the reduction of disulfide bonds of R4 by DTT indicates that disulfide bonds play an important role in self-assembly. With adhesion and coating analysis, it was found that R4 has good adhesion on different materials surfaces, which is better than Balcp-20k, while S0R4 has weak adhesion, which is only better than BSA.

PDIA6 involves the thermal stress response of razor clam, Sinonovacula constricta

Protein disulfide isomerases A6 (PDIA6), an oxidoreductase and isomerase, catalyzes the oxidation reduction and isomerization of disulfide bonds, and serves as molecular chaperone to prevent the buildup of misfolded proteins under various environmental insults. However, the role of PDIA6 in mollusks remains largely obscure, although its multifunctional protein has been reported in other species under adverse conditions. To fill this gap, we identified PDIA6 from the razor clam Sinonovacula constricta (ScPDIA6) and investigated its expression patterns in response to thermal stress. Tissue distribution showed that the mRNA transcript of ScPDIA6 was ubiquitously expressed in nine tested tissues. Temporal expression profiles by qPCR revealed that ScPDIA6 in the gill and mantle was significantly increased by hyper-thermic treatment. Further, Western blot and immunofluorescence indicated that ScPDIA6 was significantly upregulated by thermal treatment at the protein level. Additionally, the survival test demonstrated that the viability of E. coli cells expressing recombinant ScPDIA6 protein increased at 42 °C compared with empty vector. Overall, these findings suggested that ScPDIA6 may play a pivotal role in counteracting thermal stress. This study will provide valuable reference data resource for understanding the potential role of PDIA6 in mollusks.

A Combination of Native LC-MS Approaches for the Comprehensive Characterization of the Antibody-Drug Conjugate Trastuzumab Deruxtecan.

Native mass spectrometry (nMS) approaches appear attractive to complement bottom-up strategies traditionally used in biopharmaceutical industries thanks to their quite straightforward and rapid workflows, especially through online hyphenation of non-denaturing liquid chromatography (LC) to nMS. The present work provides an overview of the state-of-the-art chromatographic tools available for the detailed characterization of monoclonal antibody (mAb) formats, exemplified on the antibody-drug conjugate (ADC) trastuzumab deruxtecan (T-DXd). T-DXd was first characterized by conventional reversed phase LC (rpLC) and peptide mapping. Couplings of size exclusion chromatography (SEC), cation exchange chromatography (CEX), and hydrophobic interaction chromatography (HIC) to nMS were used to gain further insights into size, hydrophobic, and charge variants of T-DXd and its parental mAb trastuzumab, at intact and middle-up levels. SEC-nMS first offered a direct snapshot of the homogeneous conjugation of T-DXd, with an average drug-to-antibody ratio (DAR) of 8 in agreement with a conjugation on cysteines after reduction of all interchain disulfide bonds. Moreover, SEC-nMS afforded precise identification and quantification of aggregates and fragments. Middle-up level experiments performed after IdeS digestion confirmed that drug conjugation occurs in the Fab region of the mAb, as seen with rpLC. HIC separated two DAR8 species that could not be differentiated by nMS. Although middle-up HIC-nMS proved to be more informative for oxidized forms, the identification of minor variants was still difficult because of poor MS signal quality, showing how the coupling of HIC to nMS remains challenging. Lastly, middle-up CEX-nMS provided accurate determination and localization of post-translational modifications, with several acidic/basic variants within Fab and Fc regions of T-DXd that were also identified by peptide mapping. This study illustrates the strengths and drawbacks of each LC-nMS coupling. By combining SEC-, HIC-, and CEX-nMS, we were able to achieve a comprehensive characterization of T-DXd without extensive sample preparation prior to MS analysis.

Reduction pathway of glutaredoxin 1 investigated with QM/MM molecular dynamics using a neural network correction.

Glutaredoxins are small enzymes that catalyze the oxidation and reduction of protein disulfide bonds by the thiol-disulfide exchange mechanism. They have either one or two cysteines in their active site, resulting in different catalytic reaction cycles that have been investigated in many experimental studies. However, the exact mechanisms are not yet fully known, and to our knowledge, no theoretical studies have been performed to elucidate the underlying mechanism. In this study, we investigated a proposed mechanism for the reduction of the disulfide bond in the protein HMA4n by a mutated monothiol Homo sapiens glutaredoxin and the co-substrate glutathione. The catalytic cycle involves three successive thiol-disulfide exchanges that occur between the molecules. To estimate the regioselectivity of the different attacks, classical molecular dynamics simulations were performed and the trajectories analyzed regarding the sulfur-sulfur distances and the attack angles between the sulfurs. The free energy profile of each reaction was obtained with hybrid quantum mechanical/molecular mechanical metadynamics simulations. Since this required extensive phase space sampling, the semi-empirical density functional tight-binding method was used to describe the reactive cysteines. For an accurate description, we used specific reaction parameters fitted to B3LYP energies of the thiol-disulfide exchange and a machine learned energy correction that was trained on coupled-cluster single double perturbative triple [CCSD(T)] energies of thiol-disulfide exchanges. Our calculations show the same regiospecificity as observed in the experiment, and the obtained barrier heights are about 12 and 20 kcal/mol for the different reaction steps, which confirms the proposed pathway.

Oxidative stress–induced autonomous activation of the calcium/calmodulin-dependent kinase II involves disulfide formation in the regulatory domain

Calcium/calmodulin-dependent protein kinase II δ (CaMKIIδ) has a pivotal role in cardiac signaling. Constitutive and deleterious CaMKII "autonomous" activation is induced by oxidative stress, and the previously reported mechanism involves oxidation of methionine residues in the regulatory domain. Here, we demonstrate that covalent oxidation leads to a disulfide bond with Cys273 in the regulatory domain causing autonomous activity. Autonomous activation was induced by treating CaMKII with diamide or histamine chloramine, two thiol-oxidizing agents. Autonomy was reversed when the protein was incubated with DTT or thioredoxin to reduce disulfide bonds. Tryptic mapping of the activated CaMKII revealed formation of a disulfide between Cys273 and Cys290 in the regulatory domain. We determined the apparent pKa of those Cys and found that Cys273 had a low pKa while that of Cys290 was elevated. The low pKa of Cys273 facilitates oxidation of its thiol to the sulfenic acid at physiological pH. The reactive sulfenic acid then attacks the thiol of Cys290 to form the disulfide. The previously reported CaMKII mutant in which methionine residues 281 and 282 were mutated to valine (MMVV) protects mice and flies from cardiac decompensation induced by oxidative stress. Our initial hypothesis was that the MMVV mutant underwent a conformational change that prevented disulfide formation and autonomous activation. However, we found that the thiol-oxidizing agents induced autonomy in the MMVV mutant and that the mutant undergoes rapid degradation by the cell, potentially preventing accumulation of the injurious autonomous form. Together, our results highlight additional mechanistic details of CaMKII autonomous activation.

ЦИСТАМИН И ЦИСТИН ПОДАВЛЯЮТ ТРАНСПОРТ Na+ В ЭПИТЕЛИИ КОЖИ ЛЯГУШКИ

Amphibian skin and other isolated epithelial systems are classical model objects for studying the mechanisms of transepithelial ion transport. Na+ transport in osmoregulatory epithelium is a complex, multicomponent system, which involves Na+ transporting proteins and signaling cascades localized in various cell membranes. The protein components of this system may be a target for oxidative stress. At the same time, the effect of oxidizing and reducing agents on Na+ transport in native epithelial systems, such as frog skin epithelium, practically has not been studied. Using voltage-clamp technique the effect of disulfide-containing oxidizing agents cystine and cystamine on Na+ transport in frog skin was investigated. It has been shown for the first time that the treatment of the skin of the Rana temporaria frog with cystamine or cystine at a concentration of 10 μg/ml suppresses the transepithelial Na+ transport. It was also found that the preincubation with dithiothreitol, which reduces disulfide bonds in proteins, prevents the inhibitory effect of cystamine and cystine. The obtained results indicate that Na+ transport in the frog skin is sensitive to oxidative stress and is modulated by disulfide-containing oxidizing agents, such as cystamine or cystine, and also that the effect of cystamine and cystine on Na+ transport in the frog skin epithelium is mediated by their interaction with functionally important cysteine residues of Na+-transporting proteins.

Injectable redox albumin-based hydrogel with in-situ loaded dihydromyricetin

Albumin is widely used in clinics due to its demonstrated biological safety and functional flexibility. Hydrogels derived from natural albumin possess high moisture retention ability and good biodegradability, making albumin ideal biomaterials compared with synthetic polymers. Herein, by reducing disulfide bonds in bovine serum albumin molecules with glutathione and re-oxidizing the free thiols using dimethyl sulfoxide (DMSO) as additional oxidant, three-dimensional network was assembled, leading to the formation of hydrogel. Meanwhile, DMSO is also an excellent solvent for many drugs, and the hydrophobic drug dihydromyricetin (DMY) can be well dissolved in DMSO. During the crosslinking reaction, DMSO participated in fabricating a porous albumin hydrogel network. At the same time, increased loading of DMY and sustained release of DMY were achieved, improving bioavailability of hydrophobic drug DMY. Rheological test and cytotoxicity assay proved excellent elasticity and biocompatibility of the hydrogel. Self-healing property and narrow-needle injection provided potential application of the hydrogel as biomedical materials. This method for formation hydrogels and in situ loading of drugs may expand to preparing other drug loaded hydrogels and find wide applications.

Self-assembled albumin nanoparticles for redox responsive release of curcumin

The synthesis of stimuli-responsive protein-based nanocarriers often involves multi-step processing or use of toxic crosslinkers, which impede their wide-spread utilization as drug delivery systems. In this study, we report a facile, self-assembly based method for the synthesis of redox responsive albumin nanocarriers (<150 nm) without the use of crosslinkers. Curcumin (CUR) was encapsulated into these self-assembled albumin nanoparticles with 83.22% encapsulation efficiency and 8.33% loading capacity. These CUR loaded redox responsive albumin nanoparticles displayed glutathione (GSH) triggered CUR release due to the reduction of disulfide bonds in the protein structure. The release studies show that in the presence of GSH, 61% of the CUR is released, whereas in GSH-free environment only 18% of the drug is released within the first 12 h. In vitro studies performed on triple negative human breast cancer cells (MDA-MB-231) show that encapsulation of CUR provided enhanced cytotoxicity compared to free CUR. Furthermore, studies on the mitochondrial structure of these cells confirm that the drug loaded nanoparticles lead to the changes in mitochondria stress conditions by reacting with the GSH in the cells.

Relevance of lipoproteins, membranes, and extracellular vesicles in understanding C-reactive protein biochemical structure and biological activities.

Early purification protocols for C-reactive protein (CRP) often involved co-isolation of lipoproteins, primarily very low-density lipoproteins (VLDLs). The interaction with lipid particles was initially attributed to CRP’s calcium-dependent binding affinity for its primary ligand—phosphocholine—the predominant hydrophilic head group expressed on phospholipids of most lipoprotein particles. Later, CRP was shown to additionally express binding affinity for apolipoprotein B (apo B), a predominant apolipoprotein of both VLDL and LDL particles. Apo B interaction with CRP was shown to be mediated by a cationic peptide sequence in apo B. Optimal apo B binding required CRP to be surface immobilized or aggregated, treatments now known to structurally change CRP from its serum soluble pentamer isoform (i.e., pCRP) into its poorly soluble, modified, monomeric isoform (i.e., mCRP). Other cationic ligands have been described for CRP which affect complement activation, histone bioactivities, and interactions with membranes. mCRP, but not pCRP, binds cholesterol and activates signaling pathways that activate pro-inflammatory bioactivities long associated with CRP as a biomarker. Hence, a key step to express CRP’s biofunctions is its conversion into its mCRP isoform. Conversion occurs when (1) pCRP binds to a membrane surface expressed ligand (often phosphocholine); (2) biochemical forces associated with binding cause relaxation/partial dissociation of secondary and tertiary structures into a swollen membrane bound intermediate (described as mCRPm or pCRP*); (3) further structural relaxation which leads to total, irreversible dissociation of the pentamer into mCRP and expression of a cholesterol/multi-ligand binding sequence that extends into the subunit core; (4) reduction of the CRP subunit intrachain disulfide bond which enhances CRP’s binding accessibility for various ligands and activates acute phase proinflammatory responses. Taken together, the biofunctions of CRP involve both lipid and protein interactions and a conformational rearrangement of higher order structure that affects its role as a mediator of inflammatory responses.

Enzymatic basis of the Fc-selective intra-chain disulfide reduction and free thiol content variability in an antibody produced in Escherichia coli

BackgroundEscherichia coli (E. coli) is a promising host for production of recombinant proteins (including antibodies and antibody fragments) that don’t require complex post-translational modifications such as glycosylation. During manufacturing-scale production of a one-armed antibody in E. coli (periplasmic production), variability in the degree of reduction of the antibody’s disulfide bonds was observed. This resulted in variability in the free thiol content, a potential critical product quality attribute. This work was initiated to understand and prevent the variability in the total free thiol content during manufacturing.ResultsIn this study, we found that the reduction in antibody’s disulfide bonds was observed to occur during homogenization and the ensuing homogenate hold step where in the antibody is exposed to redox enzymes and small molecule reductants present in homogenate. Variability in the downstream processing time between the start of homogenization and end of the homogenate hold step resulted in variability in the degree of antibody disulfide bond reduction and free thiol content. The disulfide bond reduction in the homogenate is catalyzed by the enzyme disulfide bond isomerase C (DsbC) and is highly site-specific and occurred predominantly in the intra-chain disulfide bonds present in the Fc CH2 region. Our results also imply that lack of glycans in E. coli produced antibodies may facilitate DsbC accessibility to the disulfide bond in the Fc CH2 region, resulting in its reduction.ConclusionsDuring E. coli antibody manufacturing processes, downstream processing steps such as homogenization and subsequent processing of the homogenate can impact degree of disulfide bond reduction in the antibody and consequently product quality attributes such as total free thiol content. Duration of the homogenate hold step should be minimized as much as possible to prevent disulfide bond reduction and free thiol formation. Other approaches such as reducing homogenate temperature, adding flocculants prior to homogenization, using enzyme inhibitors, or modulating redox environments in the homogenate should be considered to prevent antibody disulfide bond reduction during homogenization and homogenate processing steps in E. coli antibody manufacturing processes.

Switching positions: Assessing the dynamics of conjugational heterogeneity in antibody-drug conjugates using CE-SDS.

Antibody-drug conjugates (ADCs) are a prospective class of new oncology therapeutics with the ability to deliver a cytotoxic drug to a targeted location. The concept appears simple, but ADCs are highly complex due to their intrinsic heterogeneity. Randomly conjugated ADCs, for instance, are composed of conjugated species carrying between 0 and 8 linker-drug molecules, with several positional isomers that vary in drug distribution across the antibody. The drug load, expressed as drug-to-antibody ratio (DAR), is a critical quality attribute and should be well controlled, together with the distribution of drug molecules. Here, the impact of the duration of disulfide bond reduction on the DAR was investigated by quantitating the (isomeric) DAR species in ADCs produced with varying reduction times. Although hydrophobic interaction chromatography showed a constant DAR value as a function of reduction time, data obtained by non-reducing CE-SDS revealed an unexpected dynamic in the positional conjugated isomers. The insights obtained have improved our understanding of the correlation between the disulfide bond reduction, an important step in the manufacturing of a cysteine-conjugated ADC, and the conjugational heterogeneity.

Effects of Mucin and DNA Concentrations in Airway Mucus on Pseudomonas aeruginosa Biofilm Recalcitrance.

ABSTRACTThe pathological properties of airway mucus in cystic fibrosis (CF) are dictated by mucus concentration and composition, with mucins and DNA being responsible for mucus viscoelastic properties. As CF pulmonary disease progresses, the concentrations of mucins and DNA increase and are associated with increased mucus viscoelasticity and decreased transport. Similarly, the biophysical properties of bacterial biofilms are heavily influenced by the composition of their extracellular polymeric substances (EPS). While the roles of polymer concentration and composition in mucus and biofilm mechanical properties have been evaluated independently, the relationship between mucus concentration and composition and the biophysical properties of biofilms grown therein remains unknown. Pseudomonas aeruginosa biofilms were grown in airway mucus as a function of overall concentration and DNA concentration to mimic healthy, and CF pathophysiology and biophysical properties were evaluated with macro- and microrheology. Biofilms were also characterized after exposure to DNase or DTT to examine the effects of DNA and mucin degradation, respectively. Identifying critical targets in biofilms for disrupting mechanical stability in highly concentrated mucus may lead to the development of efficacious biofilm therapies and ultimately improve CF patient outcomes. Overall mucus concentration was the predominant contributor to biofilm viscoelasticity and both DNA degradation and mucin reduction resulted in compromised biofilm mechanical strength.IMPORTANCE Pathological mucus in cystic fibrosis (CF) is highly concentrated and insufficiently cleared from the airway, causing chronic inflammation and infection. Pseudomonas aeruginosa establishes chronic infection in the form of biofilms within mucus, and this study determined that biofilms formed in more concentrated mucus were more robust and less susceptible to mechanical and chemical challenges compared to biofilms grown in lower concentrated mucus. Neither DNA degradation nor disulfide bond reduction was sufficient to fully degrade biofilms. Mucus rehydration should remain a priority for treating CF pulmonary disease with concomitant multimechanistic biofilm degradation agents and antibiotics to clear chronic infection.

Antibacterial responses and functional characterization of the interferon gamma inducible lysosomal thiol reductase (GILT) protein in Chinese mitten crab (Eriocheir sinensis)

The inducible reductase of interferon gamma (IFN- γ), IFN-γ-induced lysosomal thiol reductase (GILT) is important in antiviral immunity, but its mechanism in invertebrate antimicrobial immunity is unclear. We determined that GILT protein was involved in the antibacterial immunity of Chinese mitten crab (Eriocheir sinensis). GILT protein was highly expressed in crab hemocytes and was significantly upregulated 6 h after bacterial stimulation. Recombinant E. sinensis GILT (rEsGILT) contained a CXXS active site that catalyzed disulfide bond reduction. Vibrio parahaemolyticus and Staphylococcus aureus were bound through interaction with peptidoglycan and lipopolysaccharide, respectively, and bacterial agglutination and clearance in the crabs was markedly promoted. Nevertheless, EsGILT exhibited no direct antibacterial or bactericidal activity. EsGILT also promoted crab hemocyte phagocytosis and played an anti-bacterial role, and inhibited hemocyte apoptosis. In summary, EsGILT promoted bacterial agglutination, clearance, and phagocytosis by recognizing and agglutinating pathogenic microorganisms and reduced the apoptosis level, indirectly participating in antibacterial reactions.

Resource utilization of pig hair to prepare low-cost adsorbents with high density of sulfhydryl for enhanced and trace level removal of aqueous Hg(II)

Pig hair (PH), a keratinous waste, was modified by ammonium thioglycolate in a ball milling to promote its performance of Hg(II) sequestration. The ball milling broke the hydrophobic cuticle sheath and enhanced the reduction of disulfide bond, which increased the sulfydryl content of the modified PH (BTPH) from 0.07 to 11.05 μmol/g. BTPH exhibited a significantly higher capture capacity of Hg(II) (415.4 mg/g) than PH (3.1 mg/g), as well as the commercial activated carbon (219.7 mg/g), and persisted its performance over a wide range of solution pH. Meanwhile, BTPH with a distribution coefficient of 5.703 × 105 mL/g could selectively capture Hg(II) from the water with the coexisting metal ions such as Mg(II), Cd(II) and Pb(II). Moreover, the low-cost BTPH could reduce the Hg(II) from 1.0 mg/L to well below the limit of drinkable water (2 μg/L) in real-world samples. Density functional theory (DFT) calculations and state-of-the-art characterizations illustrated that the binding of Hg(II) to sulfydryl groups was the main adsorption mechanism. Notably, BTPH decreased the mercury content of water spinaches from 24.1 to 0.50 mg/kg and thereby significantly reduced the phytotoxicity of Hg(II). This work therefore provides a sustainable way to utilize keratinous wastes for the remediation of aqueous Hg(II).

Thiol-disulfide oxidoreductase PDI1;1 regulates actin structures in Oryza sativa root cells.

The polarized and dynamic actin cytoskeleton is essential for root cell growth. Here, we report the key role of thiol-disulfide oxidoreductase PDI1;1 in actin structures. Microscopic analyses revealed that after Oryza sativa roots were exposed to H2 O2 , both actin and PDI1;1 were depolarized and arranged in a meshwork. In H2 O2 -exposed cells, actin formed intermolecularly disulfide-bonded high-molecular-weight structures, which were thiol-trapped by PDI1;1. Recombinant PDI1;1 exhibited the ability to recognize actin in an in vitro binding assay. During recovery from H2 O2 exposure, the amount of disulfide-bonded high-molecular-weight structures of actin decreased over time, but deficiency of PDI1;1 inhibited the decrease. These results suggest a PDI1;1-dependent pathway that reduces disulfide bonds in high-molecular-weight structures of actin, thus promoting their degradation.

Superfast desulfurization for protein chemical synthesis and modification

Superfast desulfurization for protein chemical synthesis and modification

Functional core/shell PMMA/P(MMA‐co‐PDSECAE)‐SH particles with thiol groups in the shell and their adsorption of heavy metal ions

AbstractThe relationships between morphology and properties are important for functional polymer particles. In this work, stable core/shell PMMA/P(MMA‐co‐PDSECAE) (C/S) latexes with uniform particle size are first prepared through semi‐continuous soap‐free seeded emulsion polymerization of methyl methacrylate (MMA) and 2‐(((2‐[pyridin‐2‐yldisulfanyl]ethoxy)carbonyl)amino) ethyl acrylate (PDSECAE), and thiol‐functionalized core/shell PMMA/P(MMA‐co‐PDSECAE)‐SH (C/S‐SH) submicron particles (SPs) are then obtained by reduction of disulfide bonds with dithiothreitol (DTT). Adsorption behaviors of heavy metal ions on the SPs are investigated. Results show that, the C/S‐SH SPs, with more thiol groups distributed closer to the surface, have nearly twice the Pb2+ adsorption capacity of the homogeneous SPs prepared via batch emulsion polymerization. The C/S‐SH SPs show strong selective adsorption capacities toward Hg2+ and Pb2+, and maximum adsorption capacities reach 0.477 and 0.303 mmol g−1, respectively. Due to their high selectivity, adsorption capacity, specific surface area and simple preparation process, these core/shell SPs have promising applications in water treatment.