Search

Cell‐penetrating peptides (CPPs) have been explored as versatile tools to transport various molecules into cells. The uptake mechanism of CPPs is still not clearly understood and most probably depends on several factors like the nature of the CPP itself, the attached cargo, the investigated cell system, and other experimental conditions, such as temperature and concentration. One of the first steps of internalization involves the interaction of CPPs with negatively charged molecules present at the outer layer of the cell membrane. Recently, thiol‐mediated uptake has been found to support the effective translocation of sulfhydryl‐bearing substances that would actually not be cell‐permeable. Within this work, we aimed to understand the relevance of thiol reactivity for the uptake mechanism of cysteine‐containing CPPs that we have developed previously in our group. Therefore, we compared the two peptides, sC18‐Cys and CaaX‐1, in their single reduced and dimeric disulfide versions. Cytotoxicity, intracellular accumulation, and impact on the internalization process of the disulfides were investigated in HeLa cells. Both disulfide CPPs demonstrated significantly stronger cytotoxic effects and membrane activity compared with their reduced counterparts. Notably, thiol‐mediated uptake could be excluded as a main driver for translocation, showing that peptides like CaaX‐1 are most likely taken up by other mechanisms. [ABSTRACT FROM AUTHOR]

Functionalizing single‐walled carbon nanotubes (SWCNTs) in a robust way that does not affect the sp2 carbon framework is a considerable research challenge. Here we describe how triiodide salts of positively charged macrocycles can be used not only to functionalize SWCNTs from the outside, but simultaneously from the inside. We employed disulfide exchange in aqueous solvent to maximize the solvophobic effect and therefore achieve a high degree of macrocycle immobilization. Characterization by Raman spectroscopy, EDX‐STEM and HR‐TEM clearly showed that serendipitously this wet‐chemical functionalization procedure also led to the encapsulation of polyiodide chains inside the nanotubes. The resulting three‐shell composite materials are redox‐active and experience an intriguing interplay of electrostatic, solvophobic and mechanical effects that could be of interest for applications in energy storage. [ABSTRACT FROM AUTHOR]

Die Funktionalisierung einwandiger Kohlenstoff‐Nanoröhren (SWCNTs) auf eine robuste Weise, die das sp2 Kohlenstoffgerüst nicht beeinträchtigt ist in der aktuellen Forschung eine große Herausforderung. Hier beschreiben wir, wie Triiodidsalze von positiv geladenen Makrozyklen verwendet werden können, um SWCNTs nicht nur von außen, sondern auch gleichzeitig von innen zu funktionalisieren. Wir setzten den Disulfidaustausch in wässrigem Lösungsmittel ein, um den solvophoben Effekt zu maximieren und so einen hohen Grad an Makrozyklus Immobilisierung zu erreichen. Die Charakterisierung mittels Raman‐Spektroskopie, EDX‐STEM und HR‐TEM zeigte deutlich, dass dieses nasschemische Funktionalisierungsprotokoll auch zur Verkapselung von Polyiodidketten im Inneren der Nanoröhren führte. Die resultierenden dreischaligen Kompositmaterialien sind redoxaktiv und weisen ein faszinierendes Zusammenspiel von elektrostatischen, solvophoben und mechanischen Effekten auf, was für Anwendungen in der Energiespeicherung von Interesse sein könnte. [ABSTRACT FROM AUTHOR]

A set of five vitrimers with glass transition temperatures in the range of 80–90 °C were designed to assess the effect of the network structure and disulfide concentration on their dynamic and mechanical properties, and to find the best performing system overall compared to the commercial Araldite LY1564/Aradur 3486 commercial thermoset system. Vitrimer networks were prepared by incorporating mono- and bifunctional epoxy reactive diluents and an amine chain extender into the Araldite LY1564/4-aminophenyldisulfide system. [ABSTRACT FROM AUTHOR]

Mechanical properties and reprocessing properties are of great significance to the serviceability and recyclability of energetic composites. However, the mechanical robustness of mechanical properties and dynamic adaptability related to reprocessing properties are inherent contradictions, which are difficult to optimize at the same time. This paper proposed a novel molecular strategy. Multiple hydrogen bonds derived from acyl semicarbazides could construct dense hydrogen bonding arrays, strengthening physical cross-linking networks. The zigzag structure was used to break the regular arrangement formed by the tight hydrogen bonding arrays, so as to improve the dynamic adaptability of the polymer networks. The disulfide exchange reaction further excited the polymer chains to form a new "topological entanglement", thus improving the reprocessing performance. The designed binder (D2000-ADH-SS) and nano-Al were prepared as energetic composites. Compared with the commercial binder, D2000-ADH-SS simultaneously optimized the strength and toughness of energetic composites. Due to the excellent dynamic adaptability of the binder, the tensile strength and toughness of the energetic composites still maintained the initial values, 96.69% and 92.89%, respectively, even after three hot-pressing cycles. The proposed design strategy provides ideas for the design and preparation of recyclable composites and is expected to promote the future application in energetic composites. [ABSTRACT FROM AUTHOR]

A set of five vitrimers with glass transition temperatures in the range of 80–90 °C were designed to assess the effect of the network structure and disulfide concentration on their dynamic and mechanical properties, and to find the best performing system overall compared to the commercial Araldite LY1564/Aradur 3486 commercial thermoset system. Vitrimer networks were prepared by incorporating mono- and bifunctional epoxy reactive diluents and an amine chain extender into the Araldite LY1564/4-aminophenyldisulfide system.

Thioredoxin/glutathione reductase from Schistosoma mansoni (SmTGR) is a multifunctional enzyme that catalyzes the reduction of glutathione (GSSG) and thioredoxin, as well as the deglutathionylation of peptide and non-peptide substrates. SmTGR structurally resembles known glutathione reductases (GR) and thioredoxin reductases (TrxR) but with an appended N-terminal domain that has a typical glutaredoxin (Grx) fold. Despite structural homology with known GRs, the site of glutathione reduction has frequently been reported as the Grx domain, based primarily on aerobic, steady-state kinetic measurements and x-ray crystallography. Here, we present an anaerobic characterization of a series of variant SmTGRs to establish the site of GSSG reduction as the cysteine pair most proximal to the FAD, Cys154/Cys159, equivalent to the site of GSSG reduction in GRs. Anaerobic steady-state analysis of U597C, U597S, U597C+C31S, and I592STOP SmTGR demonstrate that the Grx domain is not involved in the catalytic reduction of GSSG, as redox silencing of the C-terminus results in no modulation of the observed turnover number (∼0.025 s -1 ) and redox silencing of the Grx domain results in an increased observed turnover number (∼0.08 s -1 ). Transient-state single turnover analysis of these variants corroborates this, as the slowest rate observed titrates hyperbolically with GSSG concentration and approaches a limit that coincides with the respective steady-state turnover number for each variant. Numerical integration fitting of the transient state data can only account for the observed trends when competitive binding of the C-terminus is included, indicating that the partitioning of electrons to either substrate occurs at the Cys154/Cys159 disulfide rather than the previously proposed Cys596/Sec597 sulfide/selenide. Paradoxically, truncating the C-terminus at Ile592 results in a loss of GR activity, indicating a crucial non-redox role for the C-terminus., (Copyright © 2024. Published by Elsevier Inc.)

Epoxy (EP) vitrimer with excellent processing and recyclability is of great potential for industrial application. In this study, we prepared an EP vitrimer based on the low viscosity homogenous mixture of diglycidyl ether of bisphenol A (DGEBA) and a liquid hardener dimethyl 3,3′-dithiodipropionate (DTDP) catalyzed by 2,4,6-tris (dimethylaminomethyl) phenol (DMP30). The optimal curing condition was determined to be 110 °C × 12 h for the original EP vitrimer. Owing to the exchangeable disulfide crosslinks, rapid stress relaxation was found for the vitrimer with a relaxation time of 1.9 s at 160 °C. Moreover, the prepared EP vitrimer possessed excellent self-healing ability and recyclability, with recovery ratio of at least 94.1% during 3 times recycling. The self-healing ability, recyclability and processing performance remain excellent for the vitrimer even after incorporating 30 wt% boron nitride (BN), which can be recycled using DMF as a solvent.

Mechanical properties and reprocessing properties are of great significance to the serviceability and recyclability of energetic composites. However, the mechanical robustness of mechanical properties and dynamic adaptability related to reprocessing properties are inherent contradictions, which are difficult to optimize at the same time. This paper proposed a novel molecular strategy. Multiple hydrogen bonds derived from acyl semicarbazides could construct dense hydrogen bonding arrays, strengthening physical cross-linking networks. The zigzag structure was used to break the regular arrangement formed by the tight hydrogen bonding arrays, so as to improve the dynamic adaptability of the polymer networks. The disulfide exchange reaction further excited the polymer chains to form a new “topological entanglement”, thus improving the reprocessing performance. The designed binder (D2000-ADH-SS) and nano-Al were prepared as energetic composites. Compared with the commercial binder, D2000-ADH-SS simultaneously optimized the strength and toughness of energetic composites. Due to the excellent dynamic adaptability of the binder, the tensile strength and toughness of the energetic composites still maintained the initial values, 96.69% and 92.89%, respectively, even after three hot-pressing cycles. The proposed design strategy provides ideas for the design and preparation of recyclable composites and is expected to promote the future application in energetic composites.

It was shown for the first time that solid amines can act as catalysts for disulfide‐based dynamic combinatorial chemistry (DCC) by ball mill grinding. The mechanochemical equilibrium for the two disulfide reactions studied was reached within 1–3 h using ten different amine catalysts. This contrasts with the weeks to months to achieve solution equilibrium for most solid amine catalysts at 2 %mol mol−1 concentration in a 2 mMolar disulfide dynamic combinatorial library in a suitable solvent. The final mechanochemical equilibrium was independent of the catalyst used but varied with other ball mill grinding factors such as the presence of traces of solvent. The different efficiencies of the amines tested were discussed. [ABSTRACT FROM AUTHOR]

Thiol‐ene click reaction is widely used in polymer chemistry due to its high efficiency and regioselectivity, and disulfide exchange can endow a covalently crosslinked network with self‐healing property. In order to explore the feasibility of introducing disulfide bonds into the polymer network via thiol‐ene click reaction, first, a nine‐function ene‐terminated dendrimer (NETD) was synthesized. Then, the polysulfide dithiol (PSDT) was used as a crosslinker to form the topological polysulfide polymer network (PSPN) under UV light/sunlight. The topological PSPN exhibited outstanding self‐healing property at moderate temperature and recovery of mechanical strength. This result was achieved by the exchangeable disulfide links in the network, leading to renewal of crosslinks across the damaged surface. [ABSTRACT FROM AUTHOR]

Starting from in-house synthesized allylic monomers containing dynamic disulfide (S S) bonds, a set of vitrimeric thiol-ene elastomers were prepared and characterized with respect to their viscoelastic and tensile properties. A mixture of a disulfide-containing diallyl monomer and disulfide-free diallyl, namely diallyl phthalate, was combined stoichiometrically with a thiol crosslinker, trimethylolpropane tris (3-mercaptopropionate), to obtain soft thiol-ene thermosets with varying degrees of dynamic bond content. The materials were able to relax strain induced stress completely and rapidly at moderately high temperatures when the disulfide content was above a certain threshold, which was accurately predicted by a statistically-based network (de)crosslinking model. Relaxation of materials with disulfide content below this threshold required the activation of an additional transesterification bond exchange process by increasing the temperature. Recycling of the materials could be achieved by hot-press molding at convenient temperatures. Especially in low-disulfide containing materials, changes were observed in the material structure and properties with increasing number of reprocessing cycles. However, a significant reduction in ambient temperature creep was observed upon reducing the disulfide content. The disulfide content can be easily optimized to yield minimal loss of mechanical property and minimal creep. This work opens up new avenues for the cost-effective development of vitrimeric elastomers with minimal ambient temperature creep. [Display omitted] • Vitrimeric thiol-ene elastomers were prepared starting from diallyl precursors containing disulphide (S S) bonds. • Complete stress relaxation was achieved with reduced disulphide allyl content at moderate temperatures. • Fully recyclable materials showing minimal ambient temperature creep were obtained. [ABSTRACT FROM AUTHOR]

Efficient cellular delivery of biologically active molecules is one of the key factors that affect the discovery and development of novel drugs. The plasma membrane is the first barrier that prevents direct translocation of chemic entities, and thus obstructs their efficient intracellular delivery. Generally, hydrophilic small molecule drugs are poor permeability that reduce bioavailability and thus limit the clinic application. The cellular uptake of macromolecules and drug carriers is very inefficient without external assistance. Therefore, it is desirable to develop potent delivery systems for achieving effective intracellular delivery of chemic entities. Apart from of the types of delivery strategies, the composition of the cell membrane is critical for delivery efficiency due to the fact that cellular uptake is affected by the interaction between the chemical entity and the plasma membrane. In this review, we aimed to develop a profound understanding of the interactions between delivery systems and components of the plasma membrane. For the purpose, we attempt to present a broad overview of what delivery systems can be used to enhance the intracellular delivery of poorly permeable chemic entities, and how various delivery strategies are applied according to the components of plasma membrane.

A method for the determination of the sulfur amino acids methionine and cysteine in milk- and plant-derived protein powders and in protein-based nutritional products is described. Samples and calibration standards are prepared by acid hydrolysis (6-M HCl, 110 °C, 24 h) with 3,3'dithiopropionic acid, evaporation, and resuspension. The released methionine and cysteine mixed disulfide are quantified by direct LC/UV, without pre- or post-column chromophore/fluorophore derivatization. Since tyrosine is also released by the acid hydrolysis and elutes in the same chromatogram, it was included in all determinations. Method performance has been defined by assessments of linearity (R2 averaged > 0.999, n = 10, for 5-point plots of each analyte), intermediate precision (within-day RSD < 2%, n = 3, and day-to-day RSD < 3%, n = 3 days, for each analyte), accuracy (cystine spike recovery = 101%; measured methionine, cystine, and tyrosine concentrations were 95–111% of published concentrations), selectivity (reagent blanks and peak purity found analyte bias ≤ 2%), and quantitation limit (25 mg of methionine and tyrosine, and 50 mg of cystine, per 100 g of protein powder). The method provides a relatively simple means for the accurate and precise determination of both sulfur amino acids in protein powders and in nutritional products, and may be performed by conventional LC/UV, without specialized amino acid analysis instrumentation. [ABSTRACT FROM AUTHOR]

The occurrence of steroid hormones in water and their serious impact on human and ecosystem demand high performance materials for efficient removal of such micropollutants. Here, an affinity membrane is developed for hormone removal with regenerable binding sites. By using photodynamic disulfides as a linker, UV induced detachment of β‐CD ligands from the membrane surface is demonstrated. The macroporous base membrane is first fabricated via a polymerization induced phase separation method using 2‐hydroxyethyl methacrylate (HEMA) and ethylene dimethylacrylate (EDMA) monomers. Then the affinity membranes are prepared by immobilizing β‐CD ligands to the poly(HEMA‐co‐EDMA) base membrane through the 2‐carboxyethyl disulfide linker. The β‐CD functionalized affinity membrane shows a 30% increase of E2 hormone uptake compared with the base membrane, attributed to the formation of CD‐hormone host–guest inclusion complexes. The photodynamic disulfide linkers enable UV‐induced detachment of blocked β‐CD ligands from and reattachment of fresh β‐CD ligands to the membrane surface after each adsorption cycle, thus conferring the affinity membrane with excellent regenerative properties. It is anticipated that the use of dynamic covalent bonds for binding ligands will be of interest for developing smart affinity membranes with regenerable and readjustable surface properties. [ABSTRACT FROM AUTHOR]

Traditional crosslinked diene rubber has excellent thermal–mechanical properties and solvent resistance, yet it is incapable of being recycled via universal molding or injecting. Vitrimers, a new class of covalently crosslinked polymer networks, can be topologically rearranged with the associative exchange mechanism, endowing them with thermoplasticity. Introducing the concept of vitrimers into crosslinked networks for the recycling of rubbers is currently an attractive research topic. However, designing tailored rubber vitrimers still remains a challenge. Herein, polybutadiene (PB) vitrimers with different structures were prepared via partial epoxidation of double bonds and ring-opening esterification reactions. Their mechanical and relaxation properties were investigated. It was found that the increasing crosslinking density can increase tensile strength and activation energy for altering the network topology. The influence of side-group effects on their relaxation properties shows that an increase in the number of epoxy groups on the polybutadiene chain can increase the chance of an effective exchange of disulfide units. This work provides a simple network design which can tune vitrimer properties via altering the crosslinking density and side-group effects.

The formation of discrete macrocycles wrapped around single‐walled carbon nanotubes (SWCNTs) has recently emerged as an appealing strategy to functionalize these carbon nanomaterials and modify their properties. Here, we demonstrate that the reversible disulfide exchange reaction, which proceeds under mild conditions, can install relatively large amounts of mechanically interlocked disulfide macrocycles on the one‐dimensional nanotubes. Size‐selective functionalization of a mixture of SWCNTs of different diameters were observed, presumably arising from error correction and the presence of relatively rigid, curved π‐systems in the key building blocks. A combination of UV/Vis/NIR, Raman, photoluminescence excitation, and transient absorption spectroscopy indicated that the small (6,4)‐SWCNTs were predominantly functionalized by the small macrocycles 12, whereas the larger (6,5)‐SWCNTs were an ideal match for the larger macrocycles 22. This size selectivity, which was rationalized computationally, could prove useful for the purification of nanotube mixtures, since the disulfide macrocycles can be removed quantitatively under mild reductive conditions. [ABSTRACT FROM AUTHOR]

Efficient cellular delivery of biologically active molecules is one of the key factors that affect the discovery and development of novel drugs. The plasma membrane is the first barrier that prevents direct translocation of chemic entities, and thus obstructs their efficient intracellular delivery. Generally, hydrophilic small molecule drugs are poor permeability that reduce bioavailability and thus limit the clinic application. The cellular uptake of macromolecules and drug carriers is very inefficient without external assistance. Therefore, it is desirable to develop potent delivery systems for achieving effective intracellular delivery of chemic entities. Apart from of the types of delivery strategies, the composition of the cell membrane is critical for delivery efficiency due to the fact that cellular uptake is affected by the interaction between the chemical entity and the plasma membrane. In this review, we aimed to develop a profound understanding of the interactions between delivery systems and components of the plasma membrane. For the purpose, we attempt to present a broad overview of what delivery systems can be used to enhance the intracellular delivery of poorly permeable chemic entities, and how various delivery strategies are applied according to the components of plasma membrane. [ABSTRACT FROM AUTHOR]

Cell adhesion to extracellular matrix, mediated by integrin receptors, is crucial for cell survival. Receptor-ligand interaction involves conformational changes in the integrin by a mechanism not fully elucidated. In addition to several direct evidence that there is disulfide re-arrangement of integrins, we previously demonstrated a role for extracellular thiols and protein disulfide isomerase (PDI) in integrin-mediated functions using platelets as model system. Exploring the possible generality of this mechanism, we now show, using three different nucleated cells which depend on adhesion for survival, that non-penetrating blockers of free thiols inhibit α2β1 and α5β1 integrin-mediated adhesion and that disulfide exchange takes place in that process. Inhibiting extracellular PDI mimics thiol blocking. Transfection with WT or enzymatically inactive PDI increased their membrane expression and enhanced cell adhesion, suggesting that PDI level is a limiting factor and that the chaperone activity of the enzyme contributes to adhesion. Exogenously added PDI also enhanced adhesion, further supporting the limiting factor of the enzyme. These data indicate that: a) Dependence on ecto-sulfhydryls for integrin-mediated adhesion is not exclusive to the platelet; b) PDI is involved in integrin-mediated adhesion, catalyzing disulfide bond exchange; c) PDI enhances cell adhesion by both its oxidoreductase activity and as a chaperone. • Disulfide exchange is required for integrin-mediated adhesion of nucleated cells. • Protein disulfide isomerase (PDI) mediates disulfide exchange during cell adhesion. • The level of membrane associated PDI is a limiting factor for adhesion. • Both oxidoreductase and chaperone activity of PDI are involved in adhesion. [ABSTRACT FROM AUTHOR]

Abstract This work introduces a novel methodological approach to study both the geometry of complex protein networks and the nature of the interacting proteins. This approach is based on the high reactivity of Au+ ions on the surface of gold nanoparticles (AuNPs) towards thiols, that allows fast formation of a covalent bond between accessible protein thiols and AuNPs. In the case of the durum wheat semolina used in the exploratory studies reported here, the nature of proteins covalently bound to AuNPs is expected to be affected by both the compactness of the protein network and by the AuNPs size. Simple centrifugation procedures allowed recovery of the protein-loaded AuNPs that remained soluble, and the protein(s) covalently bound on the surface of soluble AuNP were identified by MS analysis of their proteolytic fragments. Gluten-forming proteins were found to be bound to soluble AuNPs only when detergents or chaotropes were added to the semolina/AuNPs suspension at room temperature. AuNPs-bound proteins also included gluten-forming proteins with no reported free thiols, suggesting that they are piggybacked on other thiol-containing gluten-forming proteins via disulfide bonds already present in the otherwise untreated semolina. The potential of this approach is discussed in terms of the possibility of developing a methodology suitable for further clarification of the geometrical features of protein networks, of the nature of the involved proteins, and of the type of interaction they establish, as well as any modifications of these features upon processing. Graphical abstract Unlabelled Image Highlights • Gold nanoparticles (AuNPs) were reacted with semolina suspension • Thiol-containing gluten-forming proteins were covalently bound to soluble AuNPs • Binding of gluten-forming proteins only occurred in the presence of chaotropes/detergents • Identity of bound proteins indicate protein-protein interactions occurring in untreated semolina [ABSTRACT FROM AUTHOR]

For the preparation of tailored polymers based on polysaccharides, an easy adaptable modification method was developed by introduction of a multifunctional linker into the polymer backbone. Dextran was functionalized with a thiolactone compound that can be further treated with amine resulting in ring opening and formation of a thiol. The functional thiol group emerging can be used for crosslinking or introduction of a further functional compound by disulfide formation. Here, the efficient esterification of thioparaconic acid after in-situ activation and studies about reactivity of the dextran thioparaconate obtained are discussed. The derivative was converted by aminolysis with model compound hexylamin and the thiol generated therefrom was subsequently converted with an activated functional thiol to the corresponding disulfide. The thiolactone, which protects the thiol, enables efficient esterification without side reactions and allows storage of the polysaccharide derivative at ambient conditions for years. Not only the multifunctional reactivity of the derivative but also the end product with a balanced ratio of hydrophobic and cationic moiety is appealing for biomedical application. [Display omitted] [ABSTRACT FROM AUTHOR]

ABSTRACT: A series of poly(methyl acrylates) incorporating a disulfide center was subjected to four different stimuli to cleave the SS bond, as the weakest member of the chain. Four polymers with molecular weights in the range of 25–93 kDa were synthesized via Cu‐based atom transfer radical polymerization starting from a difunctionalized disulfide‐containing initiator. In tetrahydrofuran, the labile disulfide center was cleaved directly by mechanical force generated by ultrasound irradiation, whereas in chloroform, competing cleavage was also triggered by radical species that were generated by non‐mechanical sonolysis. A reductive cleavage was observed upon treatment with reducing agent, and a clean reversal of the cleavage via oxidation could be obtained if the terminal bromide was first removed via hydrogenolysis. This modified polymer allowed studies on a fourth strategy for the cleavage of the disulfide, namely, dynamic covalent exchange reactions with a fluorine labeled small molecule. While partially based on established protocols, this comparative study underscores the versatility of the disulfide bond for applications in stimuli‐responsive and adaptive materials. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2018, 56, 1404–1411 [ABSTRACT FROM AUTHOR]

Plastidial thioredoxin (TRX)‐like2.1 proteins are atypical thioredoxins possessing a WCRKC active site signature and using glutathione for recycling. To obtain structural information supporting the peculiar catalytic mechanisms and target proteins of these TRXs, we solved the crystal structures of poplar TRX‐like2.1 in oxidized and reduced states and of mutated variants. These structures share similar folding with TRXs exhibiting the canonical WCGPC signature. Moreover, the overall conformation is not altered by reduction of the catalytic disulfide bond or in a C45S/C67S variant that formed a disulfide‐bridged dimer possibly mimicking reaction intermediates with target proteins. Modeling of the interaction of TRX‐like2.1 with both NADPH‐ and ferredoxin‐thioredoxin reductases (FTR) indicates that the presence of Arg43 and Lys44 residues likely precludes reduction by the plastidial FTR. [ABSTRACT FROM AUTHOR]

Abstract: The control of the connectivity between nodes of synthetic networks is still largely unexplored. To address this point we take advantage of a simple dynamic chemical system with two exchange levels that are mutually connected and can be activated simultaneously or sequentially. Dithioacetals and disulfides can be exchanged simultaneously under UV light in the presence of a sensitizer. Crossover reactions between both exchange processes produce a fully connected chemical network. On the other hand, the use of acid, base or UV light connects different nodes allowing network rewiring. [ABSTRACT FROM AUTHOR]

Deubiquitinating enzymes (DUBs) catalyze the cleavage of ubiquitin from target proteins. Ubiquitin is post-translationally attached to proteins and serves as an important regulatory signal for key cellular processes. In this study, novel activity-based probes to study DUBs were synthesized that comprise a ubiquitin moiety and a novel disulfide warhead at the C-terminus. These reagents can bind DUBs covalently by forming a disulfide bridge between the active-site cysteine residue and the ubiquitin-based probe. As disulfide bridges can be broken by the addition of a reducing agent, these novel ubiquitin reagents can be used to capture and subsequently release catalytically active DUBs, whereas existing capturing agents bind irreversibly. These novel reagents allow for the study of these enzymes in their active state under various conditions. [ABSTRACT FROM AUTHOR]

Human protein disulfide isomerase (hPDI) is a key redox-regulated thiol-containing protein operating as both oxidoreductase and molecular chaperone in the endoplasmic reticulum of cells. hPDI thiol-disulfide interchange reactions lead to the adoption of two distinct red/ox conformations with different substrate preferences. hPDI also displays high binding capacity for some endogenous steroid hormones including 17 beta-estradiol (E2) and thus contributes to the regulation of their intracellular concentration, storage and actions. The primary focus of this study was to investigate the impact of E2 binding on functional activity of recombinant hPDI. Then, we examined the effect of E2 binding on structural alteration of hPDI red/ox conformations and its influence on affinity and position of interaction using experimental and computational analysis. Our results revealed that interaction of one E2 per each hPDI molecule led to the inhibition of hPDI reductase activity and conformational changes in both oxidation states. Mutually, E2-binding position were also redox-regulated with higher affinity in oxidized hPDI compare to the reduced form. The importance of histidine-256 protonation states in distinct binding preferences of E2 were also demonstrated in hPDI red/ox conformations. These findings might pave the way for better understanding of the mechanisms behind the redox-dependent hormone-binding activity of hPDI. [ABSTRACT FROM AUTHOR]

Peroxiredoxin 1 is a member of the ubiquitous peroxiredoxin family of thiol peroxidases that catalyse the reduction of peroxides. In recent years eukaryotic peroxiredoxins have emerged as a critical component of cellular redox signalling, particularly in response to alterations in production of hydrogen peroxide. Peroxiredoxins are exquisitely sensitive to oxidation by hydrogen peroxide making them key peroxide sensing enzymes within cells. Evidence gathered over the last decade suggests that in addition to sensing the redox signal, peroxiredoxins have a major role in transducing this signal to downstream signalling proteins, ultimately contributing to regulation of diverse cellular processes including proliferation, differentiation and apoptosis. In this review we present the three current models for the sensing and signal transducing roles of peroxiredoxins, with a specific focus on mammalian peroxiredoxin 1. The evidence for each mechanism is discussed and areas for future work are identified. [ABSTRACT FROM AUTHOR]

The aim of the present study was to establish a novel polymeric excipient for liquid nasal dosage forms exhibiting viscosity increasing properties, improved mucoadhesion and stability towards oxidation in solution. In order to achieve this goal, 2-mercaptonicotinic acid was first coupled to l -cysteine by disulfide exchange reaction and after purification directly attached to the polymeric backbone of xanthan gum by carbodiimide mediated amide bond formation. The resulting conjugate was characterized with respect to the amount of coupled ligand, the in situ gelling behavior, mucoadhesive properties and stability towards oxidation. Furthermore, the influence of preactivated polymers on ciliary beat frequency (CBF) of porcine nasal epithelial cells was investigated. Results showed, that 252.52 ± 20.54 μmol of the ligand was attached per gram polymer. No free thiol groups could be detected on the polymeric backbone indicating entire preactivation. Rheological investigations of polymer mucus mixtures revealed a 1.7-fold and 2.5-fold enhanced mucoadhesion of entirely preactivated xanthan (Xan-Cys-MNA) compared to thiolated xanthan (Xan-Cys) and unmodified xanthan (Xan). Tensile force evaluation reported a 2.87 and 5.11-fold higher total work of adhesion (TWA) as well as a 1.63 and 2.41-fold higher maximum detachement force of Xan-Cys-MNA compared to Xan-Cys and Xan. In the presence of H 2 O 2 as an oxidizing agent Xan-Cys-MNA showed unlike Xan-Cys no increase in viscosity, indicating high stability towards oxidation. Addition of CaCl 2 to Xan-Cys-MNA solutions caused a decrease in viscosity at nevertheless higher total viscosity. Results from CBF studies proved nasal safety for the novel conjugate. According to these results, entirely preactivated thiolated xanthan gum seems to be a promising excipient for nasal dosage forms in order to improve drug bioavailability. [ABSTRACT FROM AUTHOR]