ABSTRACT
Metal-dependent enzymes are abundant and vital catalytic agents in nature. The functional versatility of metalloenzymes has made them common targets for improvement by protein engineering as well as mimicry by de novo designed sequences. In both strategies, the incorporation of non-canonical cofactors and/or non-canonical side chains has proved a useful tool. Less explored-but similarly powerful-is the utilization of non-canonical covalent modifications to the polypeptide backbone itself. Such efforts can entail either introduction of limited artificial monomers in natural chains to produce heterogeneous backbones or construction of completely abiotic oligomers that adopt defined folds. Herein, we review recent research applying artificial protein-like backbones in the construction of metalloenzyme mimics, highlighting progress as well as open questions in this emerging field.
Subject(s)
Metalloproteins , Protein Engineering , Metalloproteins/chemistry , Metalloproteins/metabolism , Protein Engineering/methods , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Enzymes/metabolism , Enzymes/chemistry , Models, MolecularABSTRACT
The 2011 discovery of the first rare earth-dependent enzyme in methylotrophic Methylobacterium extorquens AM1 prompted intensive research toward understanding the unique chemistry at play in these systems. This enzyme, an alcohol dehydrogenase (ADH), features a La3+ ion closely associated with redox-active coenzyme pyrroloquinoline quinone (PQQ) and is structurally homologous to the Ca2+-dependent ADH from the same organism. AM1 also produces a periplasmic PQQ-binding protein, PqqT, which we have now structurally characterized to 1.46-Å resolution by X-ray diffraction. This crystal structure reveals a Lys residue hydrogen-bonded to PQQ at the site analogously occupied by a Lewis acidic cation in ADH. Accordingly, we prepared K142A- and K142D-PqqT variants to assess the relevance of this site toward metal binding. Isothermal titration calorimetry experiments and titrations monitored by UV-Vis absorption and emission spectroscopies support that K142D-PqqT binds tightly (Kd = 0.6 ± 0.2 µM) to La3+ in the presence of bound PQQ and produces spectral signatures consistent with those of ADH enzymes. These spectral signatures are not observed for WT- or K142A-variants or upon addition of Ca2+ to PQQ ⸦ K142D-PqqT. Addition of benzyl alcohol to La3+-bound PQQ ⸦ K142D-PqqT (but not Ca2+-bound PQQ ⸦ K142D-PqqT, or La3+-bound PQQ ⸦ WT-PqqT) produces spectroscopic changes associated with PQQ reduction, and chemical trapping experiments reveal the production of benzaldehyde, supporting ADH activity. By creating a metal binding site that mimics native ADH enzymes, we present a rare earth-dependent artificial metalloenzyme primed for future mechanistic, biocatalytic, and biosensing applications.
Subject(s)
Methylobacterium extorquens , Methylobacterium extorquens/enzymology , Methylobacterium extorquens/metabolism , Metalloproteins/chemistry , Metalloproteins/metabolism , Alcohol Dehydrogenase/metabolism , Alcohol Dehydrogenase/chemistry , Crystallography, X-Ray , PQQ Cofactor/metabolism , PQQ Cofactor/chemistry , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Metals, Rare Earth/chemistry , Metals, Rare Earth/metabolism , Models, Molecular , Lanthanum/chemistry , Lanthanum/metabolismABSTRACT
In enzymatic catalysis, the redox site and Lewis acid are the two main roles played by metal to assist amino acids. However, the reported enzyme mimics only focus on the redox-active metal as redox site, while the redox-inert metal as Lewis acid has, to the best of our knowledge, not been studied, presenting a bottleneck of enzyme mimics construction. Based on this, a series of highly efficient MxV2O5·nH2O peroxidase mimics with vanadium as redox site and alkaline-earth metal ion (M2+) as Lewis acid are reported. Experimental results and theoretical calculations indicate the peroxidase-mimicking activity of MxV2O5·nH2O show a periodic change with the Lewis acidity (ion potential) of M2+, revealing the mechanism of redox-inert M2+ regulating electron transfer of V-O through non-covalent polarization and thus promoting H2O2 adsorbate dissociation. The biomimetic synergetic effect of redox site and Lewis acid is expected to provide an inspiration for design of enzyme mimics.
Subject(s)
Hydrogen Peroxide , Lewis Acids , Oxidation-Reduction , Lewis Acids/chemistry , Lewis Acids/metabolism , Hydrogen Peroxide/metabolism , Hydrogen Peroxide/chemistry , Vanadium/chemistry , Vanadium/metabolism , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Biomimetics , Peroxidase/metabolism , Peroxidase/chemistry , CatalysisABSTRACT
NAD(P)H: quinone oxidoreductase-1 (NQO1) plays critical roles in antioxidation and abnormally overexpresses in tumors. Developing a fast and sensitive method of monitoring NQO1 will greatly promote cancer diagnosis in clinical practice. This study introduces a transformative colorimetric detection strategy for NQO1, harnessing an innovative competitive substrate mechanism between NQO1 and a new NADH oxidase (NOX) mimic, cobalt-nitrogen-doped carbon nanozyme (CoNC). This method ingeniously exploits the differential consumption of NADH in the presence of NQO1 to modulate the generation of H2O2 from CoNC catalysis, which is then quantified through a secondary, peroxidase-mimetic cascade reaction involving Prussian blue (PB) nanoparticles. This dual-stage reaction framework not only enhances the sensitivity of NQO1 detection, achieving a limit of detection as low as 0.67 µg mL-1, but also enables the differentiation between cancerous and noncancerous cells by their enzymatic activity profiles. Moreover, CoNC exhibits exceptional catalytic efficiency, with a specific activity reaching 5.2 U mg-1, significantly outperforming existing NOX mimics. Beyond mere detection, CoNC serves a dual role, acting as both a robust mimic of cytochrome c reductase (Cyt c) and a cornerstone for enzymatic regeneration, thereby broadening the scope of its biological applications. This study not only marks a significant step forward in the bioanalytical application of nanozymes but also sets the stage for their expanded use in clinical diagnostics and therapeutic monitoring.
Subject(s)
Colorimetry , NAD(P)H Dehydrogenase (Quinone) , NADH, NADPH Oxidoreductases , NAD(P)H Dehydrogenase (Quinone)/metabolism , NAD(P)H Dehydrogenase (Quinone)/chemistry , Humans , NADH, NADPH Oxidoreductases/metabolism , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Multienzyme Complexes/metabolism , Multienzyme Complexes/chemistry , Cobalt/chemistry , Carbon/chemistry , Biomimetics , Limit of Detection , Nitrogen/chemistry , Hydrogen Peroxide/chemistry , Hydrogen Peroxide/metabolism , Ferrocyanides/chemistry , NAD/metabolism , NAD/chemistryABSTRACT
Biomimetic cytochrome P450 for chemical activation of environmental carcinogens is an efficient in vitro model for evaluating their mutagenicity and ultimately acquiring the metabolites that cannot be easily accessed by conventional routes of organic synthesis. Different kinds of mutagen derived from polyaromatic hydrocarbons (PAHs) by metalloporphyrin/oxidant model systems have been reported, but the underlying molecular mechanisms are poorly understood. Herein, we have for the first time demonstrated an effective surface-enhanced Raman scattering (SERS) protocol to study the dynamics and biomimetic metabolic behaviors of pyrene (Pyr) in the presence of various oxygen donors. Quantitative information on the relative concentration of Pyr and its metabolites in the biomimetic system can be extracted from the SERS spectra. On the basis of our results, we conclude that the oxidative metabolism of Pyr is highly influenced by the types and concentrations of oxygen donors, leading to the formation of 1-hydroxypyrene and dioxygenated products. Besides, the addition of an appropriate amount of an organic solvent can promote the formation of secondary oxidation products. These results offer valuable insights into the dynamics of PAHs metabolism and the regulation of their metabolic pathways in biomimetic activation. In comparison to traditional liquid chromatography-mass spectrometry, the present SERS approach is more suitable for high-throughput evaluation of the metabolic process and kinetics of PAHs. We anticipate that this approach will enable a more general and comprehensive tracking of metabolic dynamics and molecular mechanisms involved in the biomimetic activation of other xenobiotics, such as procarcinogens, promutagens, and drugs.
Subject(s)
Pyrenes , Spectrum Analysis, Raman , Spectrum Analysis, Raman/methods , Kinetics , Pyrenes/chemistry , Pyrenes/metabolism , Biomimetics , Polycyclic Aromatic Hydrocarbons/metabolism , Polycyclic Aromatic Hydrocarbons/chemistry , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Surface Properties , Activation, Metabolic , Cytochrome P-450 Enzyme System/metabolism , Oxidation-ReductionABSTRACT
The pathogenic fungus Aspergillus fumigatus utilizes a cyclic ferrioxamine E (FOXE) siderophore to acquire iron from the host. Biomimetic FOXE analogues were labeled with gallium-68 for molecular imaging with PET. [68Ga]Ga(III)-FOXE analogues were internalized in A. fumigatus cells via Sit1. Uptake of [68Ga]Ga(III)-FOX 2-5, the most structurally alike analogue to FOXE, was high by both A. fumigatus and bacterial Staphylococcus aureus. However, altering the ring size provoked species-specific uptake between these two microbes: ring size shortening by one methylene unit (FOX 2-4) increased uptake by A. fumigatus compared to that by S. aureus, whereas lengthening the ring (FOX 2-6 and 3-5) had the opposite effect. These results were consistent both in vitro and in vivo, including PET imaging in infection models. Overall, this study provided valuable structural insights into the specificity of siderophore uptake and, for the first time, opened up ways for selective targeting and imaging of microbial pathogens by siderophore derivatization.
Subject(s)
Aspergillosis , Aspergillus fumigatus , Gallium Radioisotopes , Positron-Emission Tomography , Siderophores , Staphylococcus aureus , Aspergillus fumigatus/metabolism , Aspergillus fumigatus/chemistry , Positron-Emission Tomography/methods , Siderophores/chemistry , Siderophores/metabolism , Animals , Staphylococcus aureus/metabolism , Aspergillosis/diagnostic imaging , Aspergillosis/microbiology , Gallium Radioisotopes/chemistry , Species Specificity , Mice , Ferric Compounds/chemistry , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Deferoxamine/chemistry , Peptides, CyclicABSTRACT
Cobalt and zinc nanoparticles from pyrolysis of cobalt-containing ZIF-67 and zinc-containing ZIF-90 exhibited potent organophosphorus hydrolase-mimicking activities for the hydrolysis of organophosphorus compounds within minutes at pH 9.0 and 25-40 °C. The resulting nanozymes could find potential applications in many areas such as chemical decontamination, environmental protection and defense of chemical weapons.
Subject(s)
Aryldialkylphosphatase , Cobalt , Zinc , Cobalt/chemistry , Zinc/chemistry , Aryldialkylphosphatase/metabolism , Aryldialkylphosphatase/chemistry , Hydrolysis , Metal Nanoparticles/chemistry , Pyrolysis , Metal-Organic Frameworks/chemistry , Organophosphorus Compounds/chemistry , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Hydrogen-Ion ConcentrationABSTRACT
Plastics, omnipresent in the environment, have become a global concern due to their durability and limited biodegradability, especially in the form of microparticles and nanoparticles. Polystyrene (PS), a key plastic type, is susceptible to fragmentation and surface alterations induced by environmental factors or industrial processes. With widespread human exposure through pollution and diverse industrial applications, understanding the physiological impact of PS, particularly in nanoparticle form (PS-NPs), is crucial. This study focuses on the interaction of PS-NPs with model blood proteins, emphasising the formation of a protein corona, and explores the subsequent contact with platelet membrane mimetics using experimental and theoretical approaches. The investigation involves αIIbß3-expressing cells and biomimetic membranes, enabling real-time and label-free nanoscale precision. By employing quartz-crystal microbalance with dissipation monitoring studies, the concentration-dependent cytotoxic effects of differently functionalised ~210â nm PS-NPs on HEK293 cells overexpressing αIIbß3 are evaluated in detail. The study unveils insights into the molecular details of PS-NP interaction with supported lipid bilayers, demonstrating that a protein corona formed in the presence of exemplary blood proteins offers protection against membrane damage, mitigating PS-NP cytotoxicity.
Subject(s)
Nanoparticles , Polystyrenes , Protein Corona , Humans , Polystyrenes/chemistry , Protein Corona/chemistry , Protein Corona/metabolism , Nanoparticles/chemistry , HEK293 Cells , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Lipid Bilayers/chemistry , Lipid Bilayers/metabolism , Platelet Glycoprotein GPIIb-IIIa Complex/metabolism , Platelet Glycoprotein GPIIb-IIIa Complex/chemistryABSTRACT
Control of phosphate capture and release is vital in environmental, biological, and pharmaceutical contexts. However, the binding of trivalent phosphate (PO4 3-) in water is exceptionally difficult due to its high hydration energy. Based on the anion coordination chemistry of phosphate, in this study, four charge-neutral tripodal hexaurea receptors (L1-L4), which were equipped with morpholine and polyethylene glycol terminal groups to enhance their solubility in water, were synthesized to enable the pH-triggered phosphate binding and release in aqueous solutions. Encouragingly, the receptors were found to bind PO4 3- anion in a 1 : 1 ratio via hydrogen bonds in 100 % water solutions, with L1 exhibiting the highest binding constant (1.2×103â M-1). These represent the first neutral anion ligands to bind phosphate in 100 % water and demonstrate the potential for phosphate capture and release in water through pH-triggered mechanisms, mimicking native phosphate binding proteins. Furthermore, L1 can also bind multiple bioavailable phosphate species, which may serve as model systems for probing and modulating phosphate homeostasis in biological and biomedical researches.
Subject(s)
Anions , Phosphates , Water , Phosphates/chemistry , Water/chemistry , Anions/chemistry , Hydrogen-Ion Concentration , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Hydrogen Bonding , Molecular Structure , Binding SitesABSTRACT
The Gal(α1-3)Gal is the terminal disaccharide unit of the α-Gal epitope [Gal(α1-3)Gal(ß1-4)GlcNAc], an exogenous antigenic determinant with several clinical implications, found in all non-primate mammals and in several dangerous pathogens, including certain protozoa and mycobacteria. Its absence in humans makes the α-Gal epitope an interesting target for several infectious diseases. Here we present the development of a macrocyclic tweezers-shaped receptor, resulting from the combination of the structural features of two predecessors belonging to the family of diaminocarbazole receptors, which exhibits binding properties in the low millimolar range toward the Gal(α1-3)Gal disaccharide of the α-Gal antigen.
Subject(s)
Disaccharides , Disaccharides/chemistry , Humans , Epitopes/chemistry , Macrocyclic Compounds/chemistry , Biomimetics , Biomimetic Materials/chemistry , Biomimetic Materials/metabolismABSTRACT
A key approach in developing green chemistry involves converting solar energy into chemical energy of biomolecules through photocatalysis. Photocatalysis can facilitate the regeneration of nicotinamide cofactors during redox processes. Nicotinamide cofactor biomimetics (NCBs) are economical substitutes for natural cofactors. Here, photocatalytic regeneration of NADH and reduced NCBs (NCBsred) using graphitic carbon nitride (g-C3N4) was developed. The process involves g-C3N4 as the photocatalyst, Cp*Rh(bpy)H2O2+ as the electron mediator, and Triethanolamine as the electron donor, facilitating the reduction of NAD+ and various oxidative NCBs (NCBsox) under light irradiation. Notably, the highest reduction yield of 48.32 % was achieved with BANA+, outperforming the natural cofactor NAD+. Electrochemical analysis reveals that the reduction efficiency and capacity of cofactors relies on their redox potentials. Additionally, a coupled photo-enzymatic catalysis system was explored for the reduction of 4-Ketoisophorone by Old Yellow Enzyme XenA. Among all the NCBsox and NAD+, the highest conversion ratio of over 99 % was obtained with BANA+. After recycled for 8 times, g-C3N4 maintained over 93.6 % catalytic efficiency. The photocatalytic cofactor regeneration showcases its outstanding performance with NAD+ as well as NCBsox. This work significantly advances the development of photocatalytic cofactor regeneration for artificial cofactors and its potential application.
Subject(s)
Biocatalysis , Oxidation-Reduction , Photochemical Processes , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Molecular Structure , NAD/chemistry , NAD/metabolism , Biomimetics , Niacinamide/chemistry , Niacinamide/metabolism , Nitrogen Compounds/chemistry , GraphiteABSTRACT
Living organisms in nature have undergone continuous evolution over billions of years, resulting in the formation of high-performance fracture-resistant biomineralized tissues such as bones and teeth to fulfill mechanical and biological functions, despite the fact that most inorganic biominerals that constitute biomineralized tissues are weak and brittle. During the long-period evolution process, nature has evolved a number of highly effective and smart strategies to design chemical compositions and structures of biomineralized tissues to enable superior properties and to adapt to surrounding environments. Most biomineralized tissues have hierarchically ordered structures consisting of very small building blocks on the nanometer scale (nanoparticles, nanofibers or nanoflakes) to reduce the inherent weaknesses and brittleness of corresponding inorganic biominerals, to prevent crack initiation and propagation, and to allow high defect tolerance. The bioinspired principles derived from biomineralized tissues are indispensable for designing and constructing high-performance biomimetic materials. In recent years, a large number of high-performance biomimetic materials have been prepared based on these bioinspired principles with a large volume of literature covering this topic. Therefore, a timely and comprehensive review on this hot topic is highly important and contributes to the future development of this rapidly evolving research field. This review article aims to be comprehensive, authoritative, and critical with wide general interest to the science community, summarizing recent advances in revealing the formation processes, composition, and structures of biomineralized tissues, providing in-depth insights into guidelines derived from biomineralized tissues for the design and construction of high-performance biomimetic materials, and discussing recent progress, current research trends, key problems, future main research directions and challenges, and future perspectives in this exciting and rapidly evolving research field.
Subject(s)
Biomimetic Materials , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Humans , Animals , Biomineralization , Bone and Bones/chemistry , Bone and Bones/metabolism , Biomimetics/methods , Tooth/chemistryABSTRACT
Diabetes is a chronic disease in which the levels of blood glucose are too high because the body does not effectively produce insulin to meet its needs or is resistant to insulin. ß Cells in human pancreatic islets produce insulin, which signals glucogen production by the liver and causes muscles and fat to uptake glucose. Progressive loss of insulin-producing ß cells is the main cause of both type 1 and type 2 diabetes. Heparan sulfate (HS) is a ubiquitous polysaccharide found at the cell surface and in the extracellular matrix (ECM) of a variety of tissues. HS binds to and assembles proteins in ECM, thus playing important roles in the integrity of ECM (particularly basement membrane), barrier function, and ECM-cell interactions. Islet HS is highly expressed by the pancreatic ß cells and critical for the survival of ß cells. Heparanase is an endoglycosidase and cleaves islet HS in the pancreas, resulting in ß-cell death and oxidative stress. Heparanase could also accelerate ß-cell death by promoting cytokine release from ECM and secretion by activated inflammatory and endothelial cells. We demonstrate that HS-mimicking glycopolymer, a potent heparanase inhibitor, improves the survival of cultured mouse pancreatic ß cells and protects HS contents under the challenge of heparanase in human pancreatic islets. Moreover, this HS-mimicking glycopolymer reduces the expression levels of cytokines (IL8, IL1ß, and TNFα) and the gene encoding Toll-like Receptor 2 (TLR2) in human pancreatic islets.
Subject(s)
Diabetes Mellitus, Type 2 , Insulin-Secreting Cells , Animals , Biomimetic Materials/metabolism , Cytokines/metabolism , Diabetes Mellitus, Type 2/metabolism , Endothelial Cells/metabolism , Glucuronidase , Heparitin Sulfate/metabolism , Insulin/metabolism , Insulin-Secreting Cells/metabolism , MiceABSTRACT
Cell membrane cloaking technique is bioinspired nanotechnology that takes advantage of naturally derived design cues for surface modification of nanoparticles. Unlike modification with synthetic materials, cell membranes can replicate complex physicochemical properties and biomimetic functions of the parent cell source. This technique indeed has the potential to greatly augment existing nanotherapeutic platforms. Here, we provide a comprehensive overview of engineered cell membrane-based nanotherapeutics for targeted drug delivery and biomedical applications and discuss the challenges and opportunities of cell membrane cloaking techniques for clinical translation.
Subject(s)
Cell Membrane/metabolism , Nanoparticles/metabolism , Pharmaceutical Preparations/metabolism , Animals , Biomimetic Materials/metabolism , Biomimetics/methods , Drug Delivery Systems/methods , Humans , Nanotechnology/methodsABSTRACT
A homotypic cancer cell membrane camouflaged zeolitic imidazolate framework (ZIF)-based nanoagent with co-loading of two inhibitors was developed, which could suppress the efflux of protons to induce intracellular acidic stress and down-regulate glutamine metabolism to reduce the energy supply. As a compensation, glycometabolism would be upregulated with simultaneous production of large amounts of lactic acid, which could in turn aggravate the acidosis and further realize a synergetic cancer treatment.
Subject(s)
Biomimetic Materials/chemistry , Glutamine/metabolism , Nanoparticles/chemistry , Zeolites/chemistry , Animals , Biomimetic Materials/metabolism , Biomimetic Materials/pharmacology , Biomimetic Materials/therapeutic use , Cell Survival/drug effects , Enzyme Inhibitors/chemistry , Humans , Imidazoles/chemistry , Lactic Acid/metabolism , MCF-7 Cells , Neoplasms/drug therapy , Sulfides/chemistry , Thiadiazoles/chemistryABSTRACT
Biomimetic mineralization based on self-assembly has made great progress, providing bottom-up strategies for the construction of new organic-inorganic hybrid materials applied in the treatment of hard tissue defects. Herein, inspired by the cooperative effects of key components in biomineralization microenvironments, a new type of biocompatible peptide scaffold based on flexibly self-assembling low-complexity protein segments (LCPSs) containing phosphate or phosphonate groups is developed. These LCPSs can retard the transformation of amorphous calcium phosphate into hydroxyapatite (HAP), leading to merged mineralization structures. Moreover, the application of phosphonated LCPS over phosphorylated LCPS can prevent hydrolysis by phosphatases that are enriched in extracellular mineralization microenvironments. After being coated on the etched tooth enamel, these LCPSs facilitate the growth of HAP to generate new enamel layers comparable to the natural layers and mitigate the adhesion of Streptococcus mutans. In addition, they can effectively stimulate the differentiation pathways of osteoblasts. These results shed light on the potential biomedical applications of two LCPSs in hard tissue repair.
Subject(s)
Biomimetic Materials/metabolism , Calcium Phosphates/metabolism , Dental Enamel/metabolism , Durapatite/metabolism , Organophosphonates/metabolism , Phosphates/metabolism , Biomimetics/methods , Biomineralization , HumansABSTRACT
Clinical implementation of endochondral bone regeneration (EBR) would benefit from the engineering of devitalized cartilaginous constructs of allogeneic origins. Nevertheless, development of effective devitalization strategies that preserves extracellular matrix (ECM) is still challenging. The aim of this study is to investigate EBR induced by devitalized, soft callus-mimetic spheroids. To challenge the translatability of this approach, the constructs are generated using an allogeneic cell source. Neo-bone formation is evaluated in an immunocompetent rat model, subcutaneously and in a critical size femur defect. Living spheroids are used as controls. Also, the effect of spheroid maturation towards hypertrophy is evaluated. The devitalization procedure successfully induces cell death without affecting ECM composition or bioactivity. In vivo, a larger amount of neo-bone formation is observed for the devitalized chondrogenic group both ectopically and orthotopically. In the femur defect, accelerated bone regeneration is observed in the devitalized chondrogenic group, where defect bridging is observed 4 weeks post-implantation. The authors' results show, for the first time, a dramatic increase in the rate of bone formation induced by devitalized soft callus-mimetics. These findings pave the way for the development of a new generation of allogeneic, "off-the-shelf" products for EBR, which are suitable for the treatment of every patient.
Subject(s)
Biomimetic Materials/metabolism , Bone Regeneration/physiology , Cartilage/metabolism , Tissue Engineering/methods , Tissue Scaffolds , Adult , Animals , Biomimetics/methods , Extracellular Matrix/metabolism , Female , Humans , Middle Aged , Rats , Young AdultABSTRACT
Site-specific incorporation of unnatural amino acids (UAAs) with similar incorporation efficiency to that of natural amino acids (NAAs) and low background activity is extremely valuable for efficient synthesis of proteins with diverse new chemical functions and design of various synthetic auxotrophs. However, such efficient translation systems remain largely unknown in the literature. Here, we describe engineered chimeric phenylalanine systems that dramatically increase the yield of proteins bearing UAAs, through systematic engineering of the aminoacyl-tRNA synthetase and its respective cognate tRNA. These engineered synthetase/tRNA pairs allow single-site and multi-site incorporation of UAAs with efficiencies similar to those of NAAs and high fidelity. In addition, using the evolved chimeric phenylalanine system, we construct a series of E. coli strains whose growth is strictly dependent on exogenously supplied of UAAs. We further show that synthetic auxotrophic cells can grow robustly in living mice when UAAs are supplemented.
Subject(s)
Amino Acyl-tRNA Synthetases/genetics , Directed Molecular Evolution/methods , Escherichia coli/genetics , Phenylalanine/metabolism , Protein Biosynthesis , RNA, Transfer/genetics , Amino Acids/metabolism , Amino Acids/pharmacology , Amino Acyl-tRNA Synthetases/metabolism , Animals , Base Pairing , Biomimetic Materials/metabolism , Biomimetic Materials/pharmacology , Cell Engineering , Escherichia coli/metabolism , Gene Expression , Genes, Reporter , Germ-Free Life , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , HEK293 Cells , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Transgenic , Nucleic Acid Conformation , Phenylalanine/pharmacology , Plasmids/chemistry , Plasmids/metabolism , RNA, Transfer/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolismABSTRACT
The transformation from silent to functional synapses is accompanied by the evolutionary process of human brain development and is essential to hardware implementation of the evolutionary artificial neural network but remains a challenge for mimicking silent to functional synapse activation. Here, we developed a simple approach to successfully realize activation of silent to functional synapses by controlled sulfurization of chemical vapor deposition-grown indium selenide crystals. The underlying mechanism is attributed to the migration of sulfur anions introduced by sulfurization. One of our most important findings is that the functional synaptic behaviors can be modulated by the degree of sulfurization and temperature. In addition, the essential synaptic behaviors including potentiation/depression, paired-pulse facilitation, and spike-rate-dependent plasticity are successfully implemented in the partially sulfurized functional synaptic device. The developed simple approach of introducing sulfur anions in layered selenide opens an effective new avenue to realize activation of silent synapses for application in evolutionary artificial neural networks.
Subject(s)
Biomimetic Materials/metabolism , Indium/metabolism , Neural Networks, Computer , Selenium Compounds/metabolism , Sulfur/metabolism , Synapses/metabolism , Biomimetic Materials/chemistry , Humans , Indium/chemistry , Materials Testing , Selenium Compounds/chemistry , Sulfur/chemistry , Synapses/chemistryABSTRACT
High doses of radiation can cause serious side effects and efficient radiosensitizers are urgently needed. To overcome this problem, we developed a biomimetic nanozyme system (CF) by coating pyrite (FeS2) into tumor-derived exosomes for enhanced low-dose radiotherapy (RT). CF system give FeS2 with immune escape and homologous targeting abilities. After administration, CF with both glutathione oxidase (GSH-OXD) and peroxidase (POD) activities can significantly lower the content of GSH in tumor tissues and catalyze intracellular hydrogen peroxide (H2O2) to produce a large amount of ·OH for intracellular redox homeostasis disruption and mitochondria destruction, thus reducing RT resistance. Experiments in vivo and in vitro showed that combining CF with RT (2 Gy) can provide a substantial suppression of tumor proliferation. This is the first attempt to use exosomes bionic FeS2 nanozyme for realizing low-dose RT, which broaden the prospects of nanozymes.