ABSTRACT
Pyroptosis, a new mode of regulatory cell death, holds a promising prospect in tumor therapy. The occurrence of pyroptosis can trigger the release of damage-associated molecular patterns (DAMPs) and activate the antitumor immune response. Moreover, enhancing intracellular reactive oxygen species (ROS) generation can effectively induce pyroptosis. Herein, an integrated nanoplatform (hCZAG) based on zeolitic imidazolate framework-8 (ZIF-8) with Cu2+ and Zn2+ as active nodes and glucose oxidase (GOx) loading is constructed to evoke pyroptosis. GOx can effectively elevate intracellular hydrogen peroxide (H2O2) levels to regulate the unfavorable tumor microenvironment (TME). Cu2+ can be reduced to Cu+ by endogenous overexpressed GSH and both Cu2+ and Cu+ can exert Fenton-like activity to promote ROS generation and amplify oxidative stress. In addition, the accumulation of Cu2+ leads to the aggregation of lipoylated dihydrolipoamide S-acetyltransferase (DLAT), thus resulting in cuproptosis. Notably, the outburst of ROS induced by hCZAG activates Caspase-1 proteins, leads to the cleavage of gasdermin D (GSDMD), and induces pyroptosis. Pyroptosis further elicits an adaptive immune response, leading to immunogenic cell death (ICD). This study provides effective strategies for triggering pyroptosis-mediated immunotherapy and achieving improved therapeutic effects.
Subject(s)
Glucose Oxidase , Pyroptosis , Reactive Oxygen Species , Tumor Microenvironment , Pyroptosis/drug effects , Reactive Oxygen Species/metabolism , Tumor Microenvironment/drug effects , Animals , Glucose Oxidase/metabolism , Glucose Oxidase/chemistry , Humans , Mice , Copper/chemistry , Hydrogen Peroxide/chemistry , Cell Line, Tumor , Metal-Organic Frameworks/chemistry , Metal-Organic Frameworks/pharmacology , Nanoparticles/chemistry , Immunity/drug effects , Oxidative Stress/drug effects , ImidazolesABSTRACT
ZnSeTe quantum dots (QDs) attract growing interest owing to their low threats to health and the environment. They are widely applied as emitters in displays and lighting devices. Previous findings have indicated that inorganic halides are excellent candidates for surface ligands on QDs. By incorporating inorganic halides during the synthesis process, the photoluminescence (PL) intensity and quantum yield (QY) of QDs can be significantly enhanced. However, the alteration of surface states in QDs induced by zinc halide modification and the mechanism of formation of trap-state radiative recombination processes have been less discussed. Herein, we proposed a synthesis strategy for ZnSeTe/ZnSe/ZnSeS/ZnS core/shell/shell/shell QDs modified with ZnCl2, and by comparing the morphology and elemental composition of QDs with different amounts of ZnCl2 added, we revealed the regulatory mechanism of nanocrystal growth in the presence of ZnCl2. QDs with modification of ZnCl2 exhibited broad yellow fluorescence, distinct from the intrinsic blue emission. Through spectroscopic and surface ligand analyses, we attributed this yellow emission to the intermediate state energy levels caused by the defects on the surface. Finally, we used the QDs with broad linewidth emission to fabricate a simple white-light-emitting diode (WLED). This work provided new insights into the role of inorganic ligands and the use of a single emitting material in solid-state lighting devices.
ABSTRACT
Nanocatalytic therapy (NCT) has made great achievements in tumor treatments due to its remarkable enzyme-like activities and high specificity. Nevertheless, the limited types of nanozymes and undesirable tumor microenvironments (TME) greatly weaken the therapeutic efficiency. Developing a combination therapy integrating NCT and other strategies is of great significance for optimal treatment outcomes. Herein, a AuPt-loaded Cu-doped polydopamine nanocomposite (AuPt@Cu-PDA) with multiple enzyme-like activities was rationally designed, which integrated photothermal therapy (PTT) and NCT. The peroxidase (POD)-like activity of AuPt@Cu-PDA can catalyze hydrogen peroxide (H2O2) into ·OH, and the catalase (CAT)-mimic activity can decompose H2O2 into O2 to alleviate hypoxia of TME, and O2 can be further converted into toxic ·O2- by its oxidase (OXD)-mimic activity. In addition, Cu2+ in AuPt@Cu-PDA can effectively consume GSH overexpressed in tumor cells. The boosting of reactive oxygen species (ROS) and glutathione (GSH) depletion can lead to severe oxidative stress, which can be enhanced by its excellent photothermal performance. Most importantly, the accumulation of Cu2+ can disrupt copper homeostasis, promote the aggregation of lipoylated dihydrolipoamide S-acetyltransferase (DLAT), disrupt the mitochondrial tricarboxylic acid (TCA) cycle, and finally result in cuproptosis. Collectively, photothermal and photoacoustic imaging (PTI/PAI)-guided cuproptosis-enhanced NCT/PTT can be achieved. This work may expand the application of nanozymes in synergistic therapy and provide new insights into cuproptosis-related therapeutic strategies.
Subject(s)
Apoptosis , Copper , Hydrogen Peroxide , Diagnostic Imaging , Glutathione , Photothermal Therapy , Tumor Microenvironment , Cell Line, TumorABSTRACT
Particle size might affect the inhibition behaviors of gold nanoparticles (AuNPs) on enzyme activity by influencing the density of binding sites (ρ), the association constant (Ka), the steric hindrance of enzymes by AuNPs, the binding orientations of the enzyme on AuNPs, as well as the structural changes of enzymes. In previous studies, the effects of the above-mentioned factors, which could not be ignored in the applications of enzymatic electrochemistry, were often overshadowed by the effects of surface area. In order to study the size effect on the inhibition types and inhibitory ability of enzymes by AuNPs, we investigated the inhibition behaviors of chymotrypsin (ChT) by AuNPs with three different sizes (D1-AuNCs, D3-AuNPs, and D6-AuNPs) under the same surface area concentration. The results showed that both of the inhibition types and the inhibition ability varied with the particle size of AuNPs. D1-AuNCs inhibited ChT noncompetitively, while D3/D6-AuNPs inhibited ChT competitively. Contrary to the common sense, D6-AuNPs showed a weaker inhibitory ability than D3-AuNPs. By means of zeta potential, agarose gel electrophoresis, isothermal titration calorimetry, synchronous fluorescence spectroscopy, and circular dichroism, the mechanism of the weak inhibitory ability of D6-AuNPs was found to be the standing binding orientation caused by the small curvature. This work had certain guiding significance for the biosafety of AuNPs, the development of nanoinhibitors, as well as the applications of AuNPs in enzymatic electrochemistry.
Subject(s)
Metal Nanoparticles , Gold , Binding Sites , Particle Size , Spectrometry, FluorescenceABSTRACT
Colloidal quantum dots (QDs) are a class of representative fluorescent nanomaterials with tunable, bright, and sharp fluorescent emission, with promising biomedical applications. However, their effects on biological systems are not fully elucidated. In this work, we investigated the interactions between QDs with different surface ligands and different particle sizes and α-chymotrypsin (ChT) from the thermodynamic and kinetic perspectives. Enzymatic activity experiments demonstrated that the catalytic activity of ChT was strongly inhibited by QDs coated with dihydrolipoic acid (DHLA-QDs) with noncompetitive inhibitions, whereas the QDs coated with glutathione (GSH-QDs) had weak effects. Furthermore, kinetics studies showed that different particle sizes of DHLA-QDs all had high suppressive effects on the catalytic activity of ChT. It was found that DHLA-QDs with larger particle sizes had stronger inhibition effects because more ChT molecules were bound onto the surface of QDs. This work highlights the importance of hydrophobic ligands and particle sizes of QDs, which should be considered as the primary influencing factors in the assessment of biosafety. Meanwhile, the results herein can also inspire the design of nano inhibitors.
Subject(s)
Quantum Dots , Hydrophobic and Hydrophilic Interactions , Glutathione , LigandsABSTRACT
Once nanoparticles enter into the biological milieu, nanoparticle-biomacromolecule complexes, especially the protein corona, swiftly form, which cause obvious effects on the physicochemical properties of both nanoparticles and proteins. Here, the thermodynamic parameters of the interactions between water-soluble GSH-CdSe/ZnS core/shell quantum dots (GSH-QDs) and human serum albumin (HSA) were investigated with the aid of labeling fluorescence of HSA. It was proved that the labeling fluorescence originating from a fluorophore (BDP-CN for instance) could be used to investigate the interactions between QDs and HSA. Gel electrophoresis displayed that the binding ratio between HSA and QDs was â¼2:1 by direct visualization. Fluorescence resonance energy transfer (FRET) results indicated that the distance between the QDs and the fluorophore BDP-CN in HSA was 7.2 nm, which indicated that the distance from the fluorophore to the surface of the QDs was â¼4.8 nm. Fluorescence correlation spectroscopy (FCS) results showed that HSA formed a monolayer of a protein corona with a thickness of 5.5 nm. According to the spatial structure of HSA, we could speculate that the binding site of QDs was located at the side edge (not the triangular plane) of HSA with an equilateral triangular prism. The elaboration of the thermodynamic parameters, binding ratio, and interaction orientation will highly improve the fundamental understanding of the formation of protein corona. This work has guiding significance for the exploration of the interactions between proteins and nanomaterials.
Subject(s)
Cadmium Compounds , Protein Corona , Quantum Dots , Humans , Fluorescence Resonance Energy Transfer , Protein Corona/metabolism , Serum Albumin/chemistry , Cadmium Compounds/chemistry , Spectrometry, Fluorescence , Serum Albumin, Human/metabolism , Quantum Dots/chemistry , Protein BindingABSTRACT
InP quantum dots (QDs) attract growing interest in recent years, owing to their environmental advantages upon applications in display and lighting. However, compared to Cd-based QDs and Pb-based perovskites, the synthesis of InP QDs with high optical quality is relatively more difficult. Here, we established a mid-synthetic modification approach to improve the optical properties of InP-based QDs. Tris(dimethylamino)phosphine ((DMA)3P) and indium iodide were used to prepare InP QDs with a green emission (â¼527 nm). By introducing zinc halides (ZnX2) during the mid-synthetic process, the photoluminescence quantum yield (PLQY) of the resulting InP/ZnSeS/ZnS core/shell/shell QDs was increased to >70%, and the full-width-at-half-maximum (FWHM) could be narrowed to â¼40 nm. Transmission electron microscopy clearly showed the improvement of the QDs particle size distribution after introducing ZnX2. It was speculated that ZnX2 was bound to the surface of QDs as a Z-type ligand, which not only passivated surface defects and suppressed the emission of defect states but also prevented Ostwald ripening. The InP cores were also activated by ZnX2, which made the growth of the ZnSeS shell more favorable. The photoluminescence properties started to be improved significantly only when the amount of ZnX2 exceeded 0.5 mmol. As the amount increased, more ZnX2 was distributed around the QDs to form a ligand layer, which prevented the shell precursor from crossing the ligand layer to the surface of the InP core, thus reducing the size of the InP/ZnSeS/ZnS QDs. This work revealed a new role of ZnX2 and found a method for InP QDs with high brightness and low FWHM by the mid-synthetic modification, which would inspire the synthesis of even better InP QDs.
ABSTRACT
The fibrillation process of human insulin (HI) is closely related to the therapy for type II diabetes (T2D). Due to changes in the spatial structure of HI, the fibrillation process of HI takes place in the body, which leads to a significant decrease in normal insulin levels. L-Lysine CDs with a size of around 5 nm were synthesized and used to adjust and control the fibrillation process of HI. ThT fluorescence analysis and transmission electron microscopy (TEM) characterization of the CDs showed the role of HI fibrillation from the perspective of the kinetics of HI fibrillation and regulation. Isothermal titration calorimetry (ITC) was used to explore the regulatory mechanism of CDs at all stages of HI fibrillation from the perspective of thermodynamics. Contrary to common sense, when the concentration of CDs is less than 1/50 of the HI, CDs will promote the growth of fibres, while a high concentration of CDs will inhibit the growth of fibres. The experimental results of ITC clearly prove that different concentrations of CDs will correspond to different pathways of the combination between CDs and HI. CDs have a strong ability to combine with HI during the lag time, and the degree of combination has become the main factor influencing the fibrillation process.
Subject(s)
Diabetes Mellitus, Type 2 , Quantum Dots , Humans , Lysine , Quantum Dots/chemistry , Carbon/chemistry , Diabetes Mellitus, Type 2/drug therapy , InsulinABSTRACT
A boron-dipyrromethene (BODIPY)-based fluorescent probe, BDP-CN, was synthesized in this work. It had a fluorescence emission maximum at 512 nm and a high quantum yield (48%). As evidenced by agarose gel electrophoresis and liquid chromatography-mass spectrometry, it could realize the fluorescent labeling of human serum albumin (HSA) through a thiol-cyanimide addition. Interestingly, f-HSA, defined as HSA labeled by BDP-CN, had an even higher quantum yield (77%). In addition, BDP-CN would not affect the secondary structure of HSA. Based on the successful formation of f-HSA, it was further applied to study the interactions with nanoparticles. The fluorescence quenching of f-HSA by dihydrolipoic acid-coated gold nanoclusters (DHLA-AuNCs) obeyed a dynamic mechanism, consistent with the intrinsic fluorescence quenching of HSA by DHLA-AuNCs. The association constant Ka between f-HSA and DHLA-AuNCs at 298 K was 1.5 × 105 M-1, which was the same order of magnitude as that between HSA and DHLA-AuNCs. Moreover, the interactions of f-HSA with glutathione-coated gold nanoclusters confirmed that the labeled fluorescence could replace the intrinsic fluorescence to monitor the interactions between proteins and nanoparticles. By this method, strong fluorescence ensures better stability and reproducibility, excitation at a longer wavelength reduces the damage to the proteins, and covalent conjugation with cysteine residues eliminates the inner filter effects to a great extent. Therefore, the strategy for the fluorescent labeling of HSA can be expanded to investigate a broad class of nanoparticle-protein interactions and inspire even more fluorescent labeling methods with organic dyes.
Subject(s)
Metal Nanoparticles , Serum Albumin, Human , Fluorescent Dyes , Gold/chemistry , Humans , Metal Nanoparticles/chemistry , Reproducibility of Results , Serum Albumin, Human/chemistry , Spectrometry, Fluorescence/methods , Sulfhydryl CompoundsABSTRACT
Tumor targeting therapy and photodynamic therapy are effective anti-cancer therapies. Their research progress has attracted wide attention and is one of the focuses of anti-cancer drug research and development. The design and synthesis of multifunctional organic phototheranostic agents for superior image-guided diagnosis and phototherapy play an increasingly positive role in cancer diagnosis and treatment. Herein, F16M and CyM were obtained through functional design from cyanine and F16. Physicochemical characterization and biological application results showed that CyM is a multifunctional organic biological probe, which can realize intracellular multichannel (green, yellow, red, and NIR) imaging, pH detection, and mitochondrial-targeted photodynamic therapy. As an organic phototheranostic agent, it could not only realize near-infrared imaging and photodynamic therapy in vivo and in vitro but also has excellent biocompatibility and good guiding significance for the development of multichannel imaging and mitochondrial-targeting photodynamic therapy.
Subject(s)
Biosensing Techniques , Nanoparticles , Photochemotherapy , Coloring Agents , Hydrogen-Ion Concentration , Photochemotherapy/methods , PhototherapyABSTRACT
Three-dimensional (3D) self-assembled quantum dot (QD) aerogels have attracted attention due to the combined properties of both QDs and porous materials. However, the difficulty and complexity of structural composition control limit the practical application of 3D self-assembled QDs. Hence, convenient, available and multifunction QD aerogels need to be explored to promote broader practical applications. Herein, we propose a universal and facile self-assembly method of copper indium selenium (CISe) QD aerogels based on coordination interaction between Zn2+ and carboxyl. Both experiments and Monte Carlo simulations indicate that QDs are aggregated into oligomers by Zn2+, and then the oligomers are gradually interconnected to each other to form a 3D network as the concentration of Zn2+ increases. Moreover, Zn2+-induced 3D self-assembled aerogel could be depolymerized by EDTA reversibly. In combination with CISe QDs, Zn-CISe aerogel has been successfully applied in green pollution-free environment-friendly anti-counterfeiting and encryption systems.
ABSTRACT
Nanomaterials for biological applications would inevitably encounter and interact with biomolecules, which have a profound impact on the properties, functions, and even fates of both nanomaterials and biomolecules. Among the biomolecules, lysozyme (Lys) is of great importance in defending the bacterial intruder and maintaining health. Here, the interactions between fluorescent gold nanoclusters (AuNCs) (â¼2 nm) capped with different surface ligands and Lys were thoroughly investigated. Fluorescence spectroscopic studies showed that dihydrolipoic acid (DHLA)-capped and glutathione (GSH)-capped AuNCs both quenched the intrinsic fluorescence of Lys by different quenching mechanisms. Agarose gel electrophoresis and zeta-potential assays showed that statistically one DHLA-AuNC could bind one Lys, while one GSH-AuNC could bind 3-4 Lys, providing new examples for the concept of a "protein complex". Activity assays indicated that DHLA-AuNCs heavily inhibited the enzymatic activity of Lys, while GSH-AuNCs had little effect. By synchronous fluorescence and circular dichroism spectroscopic studies, it was deduced that both AuNCs would interact with Lys by electrostatic attractions due to the distinct surface charges, and then DHLA-AuNCs would further interact with Lys by hydrophobic interactions, probably due to the hydrophobic carbon chain of DHLA and the hydrophobic side chains of amino acid residues in Lys, which was proved by the significant secondary structure changes caused by DHLA-AuNCs. Meanwhile, conformational changes induced by GSH-AuNCs with zwitterionic ligands were neglectable. Therefore, this work provided a comprehensive study of the consequences and mechanisms of the interactions between Lys and AuNCs, which was essential for the design and better use of nanomaterials as biological agents.
Subject(s)
Gold , Metal Nanoparticles , Hydrophobic and Hydrophilic Interactions , Ligands , MuramidaseABSTRACT
In the last few decades, growing numbers of fluorescent probes have been developed to detect intracellular GSH. However, the majority of probes for GSH were irreversible without monitoring the changes of intracellular GSH concentration. Therefore, recently, fluorescent probes for monitoring concentrations of GSH in real-time in living cells have come into being to address this challenge. This Perspective aimed at the development of reversible probes for GSH was organized by structural features, chemical reactions, and physicochemical properties. The reversible probes designed by a coumarin skeleton as a read-out fluorophore and the Michael addition reaction as a response mechanism accounted for most of the reported reversible probes. The performances of reversible fluorescent probes based on Michael addition could be roughly predicted by fundamental laws of kinetics and thermodynamics in physical chemistry. Essentially, the design principles included a highly reactive site for GSH, a small thermodynamic driving force, a desirable Kd of 1-10 mM, and excellent cell membrane permeability. Prospectively, the development of various mechanisms and fluorophores will be effective measures to enrich the types of reversible probes for GSH.
Subject(s)
Fluorescent Dyes/chemistry , Fluorescent Dyes/metabolism , Glutathione/metabolism , Animals , Cell Survival , Humans , Time FactorsABSTRACT
Glutathione (GSH), the most abundant nonprotein thiol found in living organisms, are involved in the etiology and progression of many human diseases including cancer. So, monitoring changes of cellular GSH levels has an important guiding significance. To date, however, majority of probes can only qualitatively detect GSH in living cells. Herein, with coumarin as the read-out fluorophore and Michael addition as the sensing mechanism, six fluorescent probes were designed and synthesized. Among them, RP-2 exhibited a reversible and extremely fast response toward GSH (half time: â¼3 s), which endowed RP-2 the capacity of real-time imaging. Among the reversible probes based on Michael addition, RP-2 had both the largest forward and reverse rate constants thus far. The reaction between RP-2 and GSH was studied in detail by density functional theory and fluorescence spectroscopy. Real-time imaging of GSH levels in living cells was achieved with a temporal resolution of seconds. To simplify the processing of images, a program was developed and validated. RP-2 was expected to serve as a new fluorescent imaging tool to understand the function of intracellular GSH in the future.
Subject(s)
Coumarins/chemistry , Fluorescent Dyes/chemistry , Glutathione/analysis , Density Functional Theory , Fluorescent Dyes/chemical synthesis , HeLa Cells , Humans , Molecular Structure , Optical Imaging , Time Factors , Tumor Cells, CulturedABSTRACT
Enzymes are an important class of biomacromolecules which catalyze many metabolic processes in living systems. Nanomaterials can be synthesized with tailored sizes as well as desired surface modifications, thus acting as promising enzyme regulators. Fluorescent gold nanoclusters (AuNCs) are a representative class of ultrasmall nanoparticles (USNPs) with sizes of â¼2 nm, smaller than most of proteins including enzymes. In this work, we chose α-chymotrypsin (ChT) and AuNCs as the model system. Activity assays and inhibition kinetics studies showed that dihydrolipoic acid (DHLA)-coated AuNCs (DHLA-AuNCs) had a high inhibitory potency (IC50 = 3.4 µM) and high inhibitory efficacy (>80%) on ChT activity through noncompetitive inhibition mechanism. In distinct contrast, glutathione (GSH)-coated AuNCs (GSH-AuNCs) had no significant inhibition effects. Fluorescence spectroscopy, agarose gel electrophoresis and circular dichroism (CD) spectroscopy were conducted to explore the underlying mechanisms. A two-step interaction model was proposed. First, both DHLA-AuNCs and GSH-AuNCs might be bound to the positively charged sites of ChT through electrostatic forces. Second, further hydrophobic interactions occurred between three tyrosine residues of ChT and the hydrophobic carbon chain of DHLA, leading to a significant structural change thus to deactivate ChT on the allosteric site. On the contrary, no such interactions occurred with GSH of zwitterionic characteristic, which explained no inhibitory effect of GSH-AuNCs on ChT. To the best of our knowledge, this is the first example of the allosteric inhibition of ChT by nano regulators. These findings provide a fundamental basis for the design and development of nano regulators.
Subject(s)
Gold , Metal Nanoparticles , Chymotrypsin , Kinetics , Thermodynamics , Thioctic Acid/analogs & derivativesABSTRACT
Considering the vital role of mitochondria in the anti-cancer mechanism of organic arsenical, the mitochondria-targeted precursor PDT-PAO-TPP was designed and synthesized. PDT-PAO-TPP, as a delocalization lipophilic cation (DLCs) which mainly accumulated in mitochondria, contributed to improve anti-cancer efficacy and selectivity towards NB4 cells. In detail, PDT-PAO-TPP inhibited the activity of PDHC resulting in the suppression of ATP synthesis and thermogenesis disorder. Additionally, the inhibition of respiratory chain complex I and IV by short-time incubation of PDT-PAO-TPP also accelerated the respiration dysfunction and continuous generation of ROS. These results led to the release of cytochrome c and activation of caspase family-dependent apoptosis. Different from the mechanism of PDT-PAO in HL-60 cells, it mainly induced the mitochondrial metabolic disturbance resulting in the intrinsic apoptosis via inhibiting the activity of PDHC in NB4 cells, which also implied that the efficacy exertion of organic arsenical was a complex process involved in many aspects of cellular function. This study systematically clarifies the anti-cancer mechanism of mitochondria-targeted organic arsenical PDT-PAO-TPP and confirms the new target PDHC of organic arsenicals, which further supports the organic arsenical as a promising anticancer drug.
Subject(s)
Antineoplastic Agents/pharmacology , Arsenicals/pharmacology , Mitochondria/drug effects , Antineoplastic Agents/chemical synthesis , Apoptosis/drug effects , Arsenicals/chemical synthesis , Cell Line, Tumor , Cell Respiration/drug effects , Cytochromes c/metabolism , Humans , Mitochondrial Membranes/metabolism , Permeability/drug effects , Pyruvate Dehydrogenase Complex/metabolism , Reactive Oxygen Species/metabolism , Thermogenesis/drug effectsABSTRACT
Cadmium (Cd)-based QDs are well studied owing to their excellent optical properties. The applications of Cd-based QDs in biomedical filed, however, is hindered by its inherent toxicity. In this study, to overcome the inherent toxicity of heavy metals, CdTe QDs were encapsulated with different shells (NAC, MPA and GSH) to reduce the leakage of Cd from the core. We studied the cytotoxicity of the three kinds of CdTe QDs on S. cerevisiae by spectroscopic, electrochemical, microscopic methods and microcalorimetric technique. Results showed that toxicity of CdTe QDs increased with the augment of QD concentration. According to the values of IC50 ((GSH-CdTe QDs (15.3 nmol/L) <â¯MPA-CdTe QDs (56.2 nmol/L) <â¯NAC-CdTe QDs (89.8 nmol/L)), the most toxic one is GSH-CdTe QDs, followed by MPA-CdTe QDs, then NAC-CdTe QDs. The coatings have contribution to their toxicity. The three kinds of QDs with the similar shape (sphere) can enter the cell by the clathrin-mediated endocytosis and lead to the different impairments. The mechanism of cytotoxicity is due to the release of Cd2+ leading elevation of intracellular reactive oxygen species (ROS), which damage mitochondria. The clathrin-mediated endocytosis is a significant factor in determining the toxicity of CdTe QDs.
Subject(s)
Cadmium Compounds/toxicity , Environmental Monitoring/methods , Quantum Dots/toxicity , Reactive Oxygen Species/metabolism , Saccharomyces cerevisiae/drug effects , Tellurium/toxicity , Cadmium Compounds/chemistry , Calorimetry , Electrochemical Techniques , Humans , Microscopy, Confocal , Microscopy, Fluorescence , Oxidation-Reduction , Quantum Dots/chemistry , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae/metabolism , Surface Properties , Tellurium/chemistryABSTRACT
Based on the potential therapeutic value in targeting metabolism for the treatment of cancer, an organic arsenical PDT-BIPA was fabricated, which exerted selective anti-cancer activity in vitro and in vivo via targeting lactate dehydrogenase A (LDHA) to remodel the metabolic pathway. In details, the precursor PDT-BIPA directly inhibited the function of LDHA and converted the glycolysis to oxidative phosphorylation causing ROS burst and mitochondrial dysfunction. PDT-BIPA also altered several gene expression, such as HIF-1α and C-myc, to support the metabolic remodeling. All these changes lead to caspase family-dependent cell apoptosis in vivo and in vitro without obvious side effect. Our results provided this organic arsenical precursor as a promising anticancer candidate and suggested metabolism as a target for cancer therapies.
Subject(s)
Arsenicals/pharmacology , Disease Progression , Lactate Dehydrogenase 5/metabolism , Neoplasms/metabolism , Neoplasms/pathology , Organic Chemicals/pharmacology , Animals , Arsenicals/chemical synthesis , Arsenicals/chemistry , Cell Death/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Respiration/drug effects , Female , Glutathione/metabolism , Humans , Ki-67 Antigen/metabolism , Lactate Dehydrogenase 5/antagonists & inhibitors , Mice, Nude , Mitochondria/drug effects , Mitochondria/metabolism , Mitochondria/ultrastructure , Organic Chemicals/chemical synthesis , Organic Chemicals/chemistry , Oxygen Consumption/drug effects , Pyruvate Dehydrogenase Complex/metabolism , Rats, Wistar , Reactive Oxygen Species/metabolism , Thioredoxins/metabolism , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: CDK6 is considered as a highly validated anticancer drug target due to its essential role in regulating cell cycle progression at G1 restriction point. Activation of CDK6 requires the phosphorylation of Thr177 on A-loop, but the structural insights of the activation mechanism remain unclear. METHODS: Herein, all-atoms molecular dynamics (MD) simulations were used to study the effects of Thr177 phosphorylation on the dynamic structure of CDK6-Vcyclin complex. RESULTS: MD results indicated that the free energy barrier of the transition from open to closed state decreased ~47.2% after Thr177 phosphorylation. Key steps along the state transition process were obtained from a cluster analysis. Binding preference of ten different inhibitors to open or closed state were also investigated through molecular docking along with MD simulations methods. CONCLUSIONS: Our results indicated that Thr177 phosphorylation increased the flexibility around the ATP-binding pocket. The transition of the ATP-binding pocket between open and closed states should be considered for understanding the binding of CDK6 inhibitors. GENERAL SIGNIFICANCE: This work could deepen the understanding of CDKs activation mechanism, and provide useful information for the discovery of new CDKs inhibitors with high affinity and specificity.
Subject(s)
Cyclin-Dependent Kinase 6/chemistry , Phosphothreonine/chemistry , Protein Kinase Inhibitors/pharmacology , Protein Processing, Post-Translational/drug effects , Adenosine Triphosphate/metabolism , Binding Sites , Cyclin-Dependent Kinase 6/antagonists & inhibitors , Cyclin-Dependent Kinase 6/metabolism , Enzyme Activation , Humans , Hydrogen Bonding , Models, Molecular , Molecular Docking Simulation , Molecular Dynamics Simulation , Phosphorylation , Protein Binding , Protein Conformation , ThermodynamicsABSTRACT
Noble metal nanoclusters (NCs) show great promise as nanoprobes for bioanalysis and cellular imaging in biological applications due to ultrasmall size, good photophysical properties, and excellent biocompatibility. In order to achieve a comprehensive understanding of possible biological implications, a series of spectroscopic measurements were conducted under different temperatures to investigate the interactions of Au NCs (â¼1.7 nm) with three model plasmatic proteins (human serum albumin (HSA), γ-globulins, and transferrin). It was found that the fluorescence quenching of HSA and γ-globulins triggered by Au NCs was due to dynamic quenching mechanism, while the fluorescence quenching of transferrin by Au NCs was a result of the formation of a Au NC-transferrin complex. The apparent association constants of the Au NCs bound to HSA, γ-globulins, and transferrin demonstrated no obvious difference. Thermodynamic studies demonstrated that the interaction between Au NCs and HSA (or γ-globulins) was driven by hydrophobic forces, while the electrostatic interactions played predominant roles in the adsorption process for transferrin. Furthermore, it was proven that Au NCs had no obvious interference in the secondary structures of these three kinds of proteins. In turn, these three proteins had a minor effect on the fluorescence intensity of Au NCs, which made fluorescent Au NCs promising in biological applications owing to their chemical and photophysical stability. In addition, by comparing the interactions of small molecules, Au NCs, and large nanomaterials with serum albumin, it was found that the binding constants were gradually increased with the increase of particle size. This work has elucidated the interaction mechanisms between nanoclusters and proteins, and shed light on a new interaction mode different from the protein corona on the surface of nanoparticles, which will highly contribute to the better design and applications of fluorescent nanoclusters.