Reduction Of Bonds Research Articles

Loop 6 and the β-hairpin flap are structural hotspots that determine cofactor specificity in the FMN-dependent family of ene-reductases.

Flavin mononucleotide (FMN)-dependent ene-reductases constitute a large family of oxidoreductases that catalyze the enantiospecific reduction of carbon-carbon double bonds. The reducing equivalents required for substrate reduction are obtained from reduced nicotinamide by hydride transfer. Most ene-reductases significantly prefer, or exclusively accept, either NADPH or NADH. Despite their usefulness in biocatalytic applications, the structural determinants for cofactor preference remain elusive. We employed the NADPH-preferring 12-oxophytodienoic acid reductase 3 from Solanum lycopersicum (SlOPR3) as a model enzyme of the ene-reductase family and applied computational and structural methods to investigate the binding specificity of the reducing coenzymes. Initial docking results indicated that the arginine triad R283, R343, and R366 residing on and close to a critical loop at the active site (loop 6) are the main contributors to NADPH binding. In contrast, NADH binds unfavorably in the opposite direction toward the β-hairpin flap within a largely hydrophobic region. Notably, the crystal structures of SlOPR3 in complex witheither NADPH4 or NADH4 corroborated these different binding modes. Molecular dynamics simulations confirmed NADH binding near the β-hairpin flap and provided structural explanations for the low binding affinity of NADH to SlOPR3. We postulate that cofactor specificity is determined by the arginine triad/loop 6 and the residue(s) controlling access to a hydrophobic cleft formed by the β-hairpin flap. Thus, NADPH preference depends on a properly positioned arginine triad, whereas granting access to the hydrophobic cleft at the β-hairpin flap favors NADH binding.

Salivary protein homology between humans and dogs: Mass spectrometry-based proteomics analysis

ObjectiveThis benchmark study aimed to investigate sex-related differences based on the identification and characterization of the salivary proteome of healthy male and female dogs using mass spectrometry (MS) technique and a homology-driven approach to analyze salivary proteins in both human and dog species utilizing protein sequence alignment technique. MethodsUnstimulated whole saliva was collected from 10 healthy Beagles. After processing the samples and determining the total protein content, in-solution protein digestion was performed involving denaturation, reduction of disulfide bonds, alkylation, and removal of interfering compounds. Samples were analyzed using LC-ESI-MS/MS. ResultsLC-ESI-MS/MS analysis identified 327 and 341 unique proteins in male and female dog saliva, respectively, of which 318 (97.25 %) in male dogs and 326 (95.60 %) in female dogs were characterized. Abundant shared proteins included albumin, BPI fold-containing family A member 2, and VWFD domain-containing protein. A notable uncharacterized protein, VWFD domain-containing protein, was among the most abundant in both sexes. Comparative analysis of 69 abundant shared proteins indicated an upregulation of CES5A, EFHD, GC, IGHM, LOC100653049, KRT10, LCP1, PGD, TPI1 in male dogs, while LOC100855593 was upregulated in female dogs. In total, 84 % (n = 229/274) and 86 % (n = 235/275) salivary proteins identified in male and female dogs, respectively, were homologous to human proteins, with an overall homology of 86 % (n = 364/423), including 15 with 100 % homology. ConclusionThe study revealed clear differences in the salivary proteomics profile of healthy male and female dogs. However, most of the salivary proteins in both male and female dogs showed homology with human salivary proteins. Clinical relevanceThe identification of unique salivary proteome profiles in male and female dogs, coupled with substantial homology to human proteins, provides promising biomarkers for health assessment, highlighting its clinical significance for diagnostics and therapeutic exploration not only in veterinary and human dentistry, but across mammalian species.

Redox Out of the Box: Catalytic Versatility Across NAD(P)H-Dependent Oxidoreductases.

The asymmetric reduction of double bonds using NAD(P)H-dependent oxidoreductases has proven to be an efficient tool for the synthesis of important chiral molecules in research and on industrial scale. These enzymes are commercially available in screening kits for the reduction of C=O (ketones), C=C (activated alkenes), or C=N bonds (imines). Recent reports, however, indicate that the ability to accommodate multiple reductase activities on distinct C=X bonds occurs in different enzyme classes, either natively or after mutagenesis. This challenges the common perception of highly selective oxidoreductases for one type of electrophilic substrate. Consideration of this underexplored potential in enzyme screenings and protein engineering campaigns may contribute to the identification of complementary biocatalytic processes for the synthesis of chiral compounds. This review will contribute to a global understanding of the promiscuous behavior of NAD(P)H-dependent oxidoreductases on C=X bond reduction and inspire future discoveries with respect to unconventional biocatalytic routes in asymmetric synthesis.

Redox mal anders: katalytische Vielseitigkeit bei NAD(P)H‐abhängigen Oxidoreduktasen

AbstractDie asymmetrische Reduktion von Doppelbindungen unter Verwendung von NAD(P)H‐abhängigen Oxidoreduktasen hat sich als effizientes Werkzeug für die Synthese wichtiger chiraler Moleküle in der Forschung und im industriellen Maßstab bewährt. Diese Enzyme sind in Screening‐Kits zur Reduktion von C=O‐ (Ketone), C=C‐ (aktivierte Alkene) oder C=N‐Bindungen (Imine) kommerziell erhältlich. Aktuelle Berichte zeigen, dass die Fähigkeit, mehrere Reduktaseaktivitäten mit verschiedenartigen C=X‐Bindungen zu beherbergen, in verschiedenen Enzymklassen auftritt – entweder nativ oder nach Mutagenese. Dies stellt die gängige Vorstellung von Oxidoreduktasen infrage, hochselektiv für eine bestimmte Art des Elektrophils zu sein. Eine Berücksichtigung dieses wenig erforschten Potentials in Screenings von Enzymen und Protein‐Engineering‐Projekten könnte dazu beitragen, ergänzende biokatalytische Prozesse für die Synthese chiraler Verbindungen zu identifizieren. Dieser Übersichtsartikel soll zum allgemeinen Verständnis des promiskuitiven Verhaltens von NAD(P)H‐abhängigen Oxidoreduktasen bei der Reduktion von C=X‐Bindungen beitragen und zukünftige Entdeckungen unkonventioneller biokatalytischer Routen in der asymmetrischen Synthese inspirieren.

Theoretical Design and Synthesis of Caged Compounds Using X-Ray-Triggered Azo Bond Cleavage.

Caged compounds are frequently used in life science research. However, the light used to activate them is commonly absorbed and scattered by biological materials, limiting their use to basic research in cells or small animals. In contrast, hard X-rays exhibit high bio-permeability due to the difficulty of interacting with biological molecules. With the main goal of developing X-ray activatable caged compounds, azo compounds are designed and synthesized with a positive charge and long π-conjugated system to increase the reaction efficiency with hydrated electrons. The azo bonds in the designed compounds are selectively cleaved by X-ray, and the fluorescent substance Diethyl Rhodamine is released. Based on the results of experiments and quantum chemical calculations, azo bond cleavage is assumed to occur via a two-step process: a two-electron reduction of the azo bond followed by N─N bond cleavage. Cellular experiments also demonstrate that the azo bonds can be cleaved intracellularly. Thus, caged compounds that can be activated by an azo bond cleavage reaction promoted by X-ray are successfully generated.

Reconnection of cysteine in reduced hair with alkylene dimaleates via thiol-Michael click chemistry.

Conventional hair permanent waving (PW) and permanent straightening processes typically involve two steps: reduction, for breaking -S-S- bond in cystine into cysteine and oxidation for -S-S- bond reconnection. However, it is known that the hair incurs damage during the oxidation step. In this work, we proposed a novel strategy to reconnect reduced disulfide bonds in hair via the thiol-Michael click reaction, by using a symmetric Michael reagent. Virgin black Chinese hair was reduced using 8% wt thioglycolic acid and employed as model hair containing a high content of broken disulfide bonds. The reduced hair was treated with 1,4-n-butylene dimaleate. Raman spectroscopy and Fourier transform infrared spectroscopy (FT-IR) were used to verify the chemical changes occurred in untreated and treated hair fibre. Single-fibre mechanical properties and thermal properties of the hair were evaluated using tensile testing and differential scanning calorimetry (DSC), respectively. The 1,4-n-butylene dimaleate could reconnect free thiol groups generated by disulfide bond reduction via thiol-Michael click reaction and significantly improve the mechanical strength of hair compared to that of the reduced hair. Secondary conformational resolution analysis of FT-IR results revealed that the content of α-helix structure could be restored after treatment with 1,4-n-butylene dimaleate. The intermolecular forces established by the newly generated C-S bonds compensate the broken disulfide bonds and enhance the fracture strength of the hair compared to that of reduced hair. Michael reagents of similar structure also showed similar performance in restoring the mechanical properties of reduced hair. Our data suggest that 1,4-n-butylene dimaleate can restore the mechanical properties of reduced hair by reconnecting reduced disulfide bonds and restoring the secondary conformation of hair keratin.

Endoplasmic Reticulum Protein 72 Regulates Integrin Mac-1 Activity to Influence Neutrophil Recruitment.

Integrins mediate the adhesion, crawling, and migration of neutrophils during vascular inflammation. Thiol exchange is important in the regulation of integrin functions. ERp72 (endoplasmic reticulum-resident protein 72) is a member of the thiol isomerase family responsible for the catalysis of disulfide rearrangement. However, the role of ERp72 in the regulation of Mac-1 (integrin αMβ2) on neutrophils remains elusive. Intravital microscopy of the cremaster microcirculation was performed to determine in vivo neutrophil movement. Static adhesion, flow chamber, and flow cytometry were used to evaluate in vitro integrin functions. Confocal fluorescent microscopy and coimmunoprecipitation were utilized to characterize the interactions between ERp72 and Mac-1 on neutrophil surface. Cell-impermeable probes and mass spectrometry were used to label reactive thiols and identify target disulfide bonds during redox exchange. Biomembrane force probe was performed to quantitatively measure the binding affinity of Mac-1. A murine model of acute lung injury induced by lipopolysaccharide was utilized to evaluate neutrophil-associated vasculopathy. ERp72-deficient neutrophils exhibited increased rolling but decreased adhesion/crawling on inflamed venules in vivo and defective static adhesion in vitro. The defect was due to defective activation of integrin Mac-1 but not LFA-1 (lymphocyte function-associated antigen-1) using blocking or epitope-specific antibodies. ERp72 interacted with Mac-1 in lipid rafts on neutrophil surface leading to the reduction of the C654-C711 disulfide bond in the αM subunit that is critical for Mac-1 activation. Recombinant ERp72, via its catalytic motifs, increased the binding affinity of Mac-1 with ICAM-1 (intercellular adhesion molecule-1) and rescued the defective adhesion of ERp72-deficient neutrophils both in vitro and in vivo. Deletion of ERp72 in the bone marrow inhibited neutrophil infiltration, ameliorated tissue damage, and increased survival during murine acute lung injury. Extracellular ERp72 regulates integrin Mac-1 activity by catalyzing disulfide rearrangement on the αM subunit and may be a novel target for the treatment of neutrophil-associated vasculopathy.

Interactions between Damaged Hair Keratin and Juglone as a Possible Restoring Agent: A Vibrational and Scanning Electron Microscopy Study.

Juglone, a quinonic compound present in walnut extracts, was proposed as a restoring agent for hair keratin treated with permanent or discoloration processes. The proposed mechanism of restoration by juglone involves the formation of a Michael adduct between the quinone and the thiol moieties of cysteine residues. To this purpose, the first part of the present paper involved the spectroscopic study of the product of the reaction between juglone and N-acetyl-L-cysteine as a model compound. IR spectroscopy and Scanning Electron Microscopy (SEM) monitored the chemical and morphological variations induced by applying juglone to hair keratin. In order to simulate the most common hair treatments (i.e., permanent and discoloration), juglone was applied to hair that had been previously treated with a reducing agent, i.e., methyl thioglycolate (MT) or with bleaching agents (based on hydrogen peroxide and persulfates) followed by sodium hydrogen sulfite. IR spectroscopy allowed us to monitor the formation of Michael adducts between juglone and cysteine residues: the Michael adducts' content was related to the cysteine content of the samples. In fact, MT and sodium hydrogen sulfite favored the reduction of the disulfide bonds and increased the content of free cysteine residues, which can react with juglone. SEM analyses confirmed the trend observed by IR spectroscopy since hair samples treated with juglone adopted a more regular hair surface and more imbricated scales, thus supporting the possible use of juglone as a restoring agent for damaged hair keratins.

Viscosity Reduction Mechanism and Rheological Properties of Ethylene-Bis-Stearamide and Crumb Rubber Modified Asphalt

This study investigates the effect of ethylene-bis-stearamide (EBS) on viscosity reducing of crumb rubber modified asphalt (CRMA) and explores the interaction mechanism via molecular dynamics simulation (MD), microscope method and rheological experiment. It is confirmed that the EBS play an important role in reducing the viscosity of ethylene-bis-stearamide and crumb rubber modified asphalt (EBS-CRMA), especially when the content is within 3%. The micro morphology and MD results showed that the EBS is coating on rubber and asphaltenes and then forming lubrication film, which is the main reason leading to the viscosity decrease of EBS-CRMA. Additionally, other factors also have significant effect in reducing the viscosity of EBS-CRMA, including the more uniform distribution of asphaltenes, the enhanced diffusion ability of saturates and resins, the reduction of hydrogen bonds and soluble substances within asphalt phase. When EBS content exceeds 3%, the influence on viscosity of EBS-CRMA is not obvious with the EBS content continuously increasing. It can be concluded from Fourier Transform Infrared (FTIR) Spectrometer and MD results that the molecular aggregation, the concentration increase of intermolecular hydrogen bonds and the diffusion ability decrease of EBS-CRMA molecular system are main reasons. Meanwhile, the coating effect of EBS and soluble substances concentration within asphalt phase are limited as the EBS content over 3%.

Redox Biology Involved in the Toxicity of Enterohemorrhagic Escherichia coli Toxin SubAB

AB5 toxins of pathogenic bacteria enter host cells and utilize the retrograde trafficking pathway to translocate to the cytoplasm and exert its pathogenesis. Cholera toxin and Shiga toxin reach the endoplasmic reticulum (ER), and the A subunit undergoes redox regulation by ER proteins to become active fragments, which pass through the ER membrane and translocate to the cytoplasm. By acting on molecular targets in the cytoplasm, the normal function of host cells are disrupted, causing diseases. ER chaperone proteins such as protein disulfide isomerase (PDI) and binding immunoglobulin protein (BiP) induce conformational changes triggered by the reduction of disulfide bonds in the A subunit. This is thought to be dependent on cysteine thiol-mediated redox regulation, but the detailed mechanism remains unclear. On the other hand, subtilase cytotoxin (SubAB), produced by enterohemorrhagic Escherichia coli (EHEC), localizes to the ER without translocating to the cytoplasm and cleaves BiP as a substrate. Therefore, it is thought that ER stress-based cytotoxicity and intestinal bleeding occur without translocating to the cytoplasm. We reported that PDI is involved in BiP cleavage through SubAB localization to the ER. Like other AB5 toxins, this indicates the involvement of redox regulation via chaperone proteins in the ER, but also suggests that SubAB does not translocate to the cytoplasm because it cleaves BiP. Although there are few reports on the redox state of ER protein thiols, it is suggested that polysulfidation, which is discussed in this symposium, may be involved.

Artemisinin: a novel chiral electrochemiluminescence luminophore-assisted enantiospecific recognition and mechanism identification.

Exploring novel electrochemiluminescence (ECL) molecules with high efficiency and good stability in aqueous solutions is crucial for achieving highly sensitive detection of analytes. However, developing chiral luminophores with efficient ECL performance is still a challenge. Herein, we first uncover that artemisinin (ART), a well-known chiral antimalarial drug, features a strong ECL emission at 726 nm with the assistance of a co-reactant potassium persulfate (K2S2O8), and an ECL efficiency of 195.3%, compared to that of standard Ru(bpy)3Cl2/K2S2O8. Mechanistic studies indicate that the strong ECL signal of ART is generated when the excited state formed by the reduction of ART peroxide bonds and combination with persulfate returns to the ground state. Significantly, we found that the ECL sensor based on chiral ART could efficiently identify and detect chiral cysteine (Cys) through ECL signals, with a lower limit of detection of 3.7 nM for l-Cys. Density functional theory calculations and scanning electrochemical microscopy technology further confirm that the disparity in the ECL signals is attributed to the different affinity between chiral ART and d/l-Cys, resulting in distinct electron transfer rates. The study demonstrates a new role of ART in ECL investigation and for the first time, achieves the development of ART for the enantioselective recognition and sensitive detection of chiral substances. This will be of vital significance for ECL and chirality research.

Helenine blocks NLRP3 activation by disrupting the NEK7-NLRP3 interaction and ameliorates inflammatory diseases.

The involvement of NLRP3 inflammasome is associated with the progress of numerous inflammatory conditions. However, there is currently no single compound used in the clinic. Search for the inhibitor of NLRP3 inflammasome from natural products is an attractive direction. The compound Helenin (Hel), which is obtained from Inula helenium L., is reported to have anti-inflammatory activities. However, the underlying molecular mechanisms and specific inflammatory signal pathway remains not well understood. This research aims to determine the impacts of Hel on NLRP3 inflammasome and the underlying mechanism involved, meanwhile also assessing its potential as a therapeutic intervention for inflammatory diseases mediated by NLRP3 overactivation. Pretreated with Hel in BMDMs (bone marrow-derived macrophages), then stimulated with NLRP3 triggers and measured the expression of active caspase-1 and interleukin 1β (IL-1β). Determination of intracellular K+ and Ca2+, ASC oligomerization and mitochondrial reactive oxygen species (mtROS) production were employed to explore the preliminary mechanism of Hel on NLRP3 activation. Subsequently, Co-immunoprecipitation was used to investigate protein-protein interaction and reduction of covalent bonds of Hel was to explore the binding mode between drugs and proteins. Finally, in vivo experiments, we utilized mouse lethal sepsis and monosodium urate(MSU)-induced peritonitis models to evaluate the effectiveness of Hel in inhibiting inflammatory diseases. The findings revealed that Hel exhibited a specific blocking effect on NLRP3, with no impact on the assembly of NLRC4 and AIM2 inflammasome. Through the analysis of mechanisms targeting key upstream factors in NLRP3 activation, Hel inhibited NLRP3-dependent ASC oligomerization but did not regulating inflammasome priming, K+ efflux, Ca2+ influx, or mitochondrial damage and mtROS. Moreover, Hel effectively interrupted the binding of NEK7-NLRP3, which was dependent on the active double C=C of the α,β-unsaturated carbonyl units in Hel. In mouse models, Hel showed promising therapeutic effects in the treatment of NLRP3 overactivation-associated diseases, including the lethal sepsis and acute systemic inflammation induced by lipopolysaccharide (LPS) and peritonitis induced by MSU. Our results indicate that Hel dependent α,β-unsaturated carbonyl units interrupt the formation of the NLRP3-NEK7 interaction, thereby blocks the inflammasome assemblage and activation. These fundings would suggest that Hel is a promising inhibitor for treating diseases driven by NLRP3 overactivation.

Photocatalytic organosulfur reagent-promoted selective mono-(deutero)hydrodechlorination

Selective partial C–Cl bond reduction represents an important strategy for the construction of valuable chlorine-containing skeletons.

Unified Models for Bond Strength and Bond-Slip Response of Deformed Rebars in Cracked Concrete

Cracks in reinforced concrete structures pose a potential risk to their long-term stability and safety, necessitating an accurate analysis of the bond failure mechanism between the reinforcement and the cracked concrete. However, currently, there is a lack of comprehensive bond prediction models for the entire cracking stage, and limited understanding of the bond slip response. To address the aforementioned issues, an improved model incorporating internal pressure, crack width, and failure modes, aiming to provide a more precise estimation of the bond strength between deformed bars embedded in concrete under various cracked cases. The proposed model demonstrates superior performance in predicting cracked cases and failure modes. The performance of the proposed model in predicting cracked cases and failure modes is superior to existing models, as demonstrated by a comparative analysis. Additionally, a comprehensive model is proposed to analyze the bond-slip behavior in cracked concrete. This model introduces a bond reduction factor for cracks, aiming to determine the critical bond length required for bond strength.

Organic Electrochemical Transistor Aptasensor for Interleukin-6 Detection.

We demonstrate an organic electrochemical transistor (OECT) biosensor for the detection of interleukin 6 (IL6), an important biomarker associated with various pathological processes, including chronic inflammation, inflammaging, cancer, and severe COVID-19 infection. The biosensor is functionalized with oligonucleotide aptamers engineered to bind specifically IL6. We developed an easy functionalization strategy based on gold nanoparticles deposited onto a poly(3,4-ethylenedioxythiophene) doped with polystyrenesulfonate (PEDOT:PSS) gate electrode for the subsequent electrodeposition of thiolated aptamers. During this functionalization step, the reduction of sulfide bonds allows for simultaneous deposition of a blocking agent. A detection range from picomolar to nanomolar concentrations for IL6 was achieved, and the selectivity of the device was assessed against Tumor Necrosis Factor (TNF), another cytokine involved in the inflammatory processes.

Manganese doped graphene oxide: Selective hydrogenation catalyst for converting 5-hydroxymethyl furfural to 5-methyl furfural

Developing a sustainable catalytic system for the production of fuels and chemicals from biomass remains a vital and challenging process. In this context, the selective hydrogenation of the biomass-derived molecule 5-hydroxymethylfurfural (HMF) to 5-methylfurfural (MF) with molecular hydrogen is one of the most important transformations but it is a difficult task because the C=O bond reduction in HMF is highly favorable than C−OH. This reaction finds applications in diverse sectors, including pharmaceuticals, and petrochemicals, etc. In this article, we report the use of manganese functionalized graphene oxide as heterogeneous catalysts for the conversion of HMF to MF, utilizing H2 as a reducing agent. The synergistic effect between graphene oxide and MnOx are responsible for the activation of C−OH in the reaction. This work unlocks the potential of the earth abundant metal-based catalyst for the selective hydrogenation of renewable biomass to chemical feedstocks.

Reduction of Triple Bond in [B12H11NCR]− Anions by Lithium Aluminum Hydride: A Novel Approach to the Synthesis of N-Monoalkylammonio-Substituted closo-Dodecaborates

By reacting nitrilium derivative of the closo-dodecaborate anion, Bu4N[B12H11N≡CR] (where R = Me, Et, nPr, iPr, p-tolyl), with lithium aluminum hydride (LiAlH4), N-alkylammonium derivatives of the closo-dodecaborate anion, and Bu4N[B12H11NH2CH2R], were obtained. The reduction reaction procedure was optimized, achieving yields close to quantitative (90–95%). The structure of the compound Bu4N[B12H11NH2CH2CH3] was determined using X-ray structural analysis. It was found that substituting lithium aluminum hydride (LiAlH4) with sodium borohydride (NaBH4) leads to the same products but only upon heating, while the reaction with LiAlH4 proceeds at room temperature.

Absorption, Distribution, Metabolism, and Excretion of [14C]BS1801, a Selenium-Containing Drug Candidate, in Rats.

BS1801 is a selenium-containing drug candidate with potential for treating liver and lung fibrosis. To fully elucidate the biotransformation of BS1801 in animals and provide sufficient preclinical drug metabolism data for human mass balance study, the metabolism of BS1801 in rats was investigated. We used radiolabeling techniques to investigate the mass balance, tissue distribution, and metabolite identification of BS1801 in Sprague-Dawley/Long-Evans rats after a single oral dose of 100 mg/kg (100 μCi/kg) [14C]BS1801: 1. The mean recovery of radioactive substances in urine and feces was 93.39% within 168 h postdose, and feces were the main excretion route. 2. Additionally, less than 1.00% of the dose was recovered from either urine or bile. 3. BS1801-related components were widely distributed throughout the body. 4. Fifteen metabolites were identified in rat plasma, urine, feces, and bile, and BS1801 was detected only in feces. 5. BS1801-M484, the methylation product obtained via a N-Se bond reduction in BS1801, was the most abundant drug-related component in plasma. The main metabolic pathways of BS1801 were reduction, amide hydrolysis, oxidation, and methylation. Overall, BS1801 was distributed throughout the body, and excreted mainly as an intact BS1801 form through feces. No differences were observed between male and female rats in distribution, metabolism, and excretion of BS1801.

Rol' vakansiy v spin-zhidkostnoy modeli Yao–Li

We consider the effect of vacancies on the low-energy excitation spectrum of a quantum spin liquid realized in the exactly solvable Yao–Lee model [H. Yao and D.-H. Lee, Phys. Rev. Lett. 107, 087205 (2011)]. Physically, vacancies can appear for different reasons (e.g., because of zero magnetic moments on the lattice, or the presence of nonmagnetic impurities, or a random reduction of local bonds of magnetic moments with the remaining lattice). It is shown numerically that the finite density of random vacancies in this model leads to the accumulation of states near zero energy, which can be detected from the change of the behavior of heat capacity at low temperatures. Moreover, it is shown that the low-energy modes are localized more strongly than remaining eigenmodes. This effect is illustrated using the inverse participation ratio (IPR). In the case of time reversal symmetry breaking (e.g., due to the presence of a magnetic field), a gap is opened in the fermion spectrum of the model, and vacancy-induced localized states appear. The energies of these states depend on the structure of the interactions responsible for the time inversion symmetry breaking.

Heterogeneity in Disulfide Bond Reduction in IgG1 Antibodies Is Governed by Solvent Accessibility of the Cysteines.

We studied unpaired cysteine levels and disulfide bond susceptibility in four different γ-immunoglobulin antibodies using liquid chromatography-mass spectrometry. Our choice of differential alkylating agents ensures that the differential peaks are non-overlapping, thus allowing us to accurately quantify free cysteine levels. For each cysteine residue, we observed no more than 5% to be unpaired, and the free cysteine levels across antibodies were slightly higher in those containing lambda light chains. Interchain and hinge residues were highly susceptible to reducing stresses and showed a 100-1000-fold higher rate of reduction compared to intrachain cysteines. Estimations of the solvent-accessible surface for individual cysteines in IgG1, using an implicit all-atom molecular dynamics simulation, show that interchain and hinge cysteines have >1000-fold higher solvent accessibility compared to intrachain cysteines. Further analyses show that solvent accessibility and the rate of reduction are linearly correlated. Our work clearly establishes the fact that a cysteine's accessibility to the surrounding solvent is one of the primary determinants of its disulfide bond stability.