RESUMEN
Owing to the unique catalytic, optical and magnetic properties, lanthanides (Ln) as multicomponent biomarkers, are widely used in the field of optical sensing, mass spectrometry and magnetic resonance imaging. As ligands, DNA molecules have good biocompatibility, high stability, cost efficiency, programmability and biodegradability. Based on the coordination-driven self-assembly between Ln ions (Ln3+ ) and DNA molecules, a multifunctional Ln3+ -DNA hybrid coordination polymers (CPs) were synthesized. Not only a series of different Ln3+ (single Ln3+ ) and DNA hybrid CPs were synthesized, but one hybrid CP contains two kinds of Ln3+ was obtained. Besides, the synthetic CPs in cell fluorescence imaging and miRNA sensing also exhibited high performance. This work provides a novel idea for the synthesis of DNA based nanomaterials, which is promising for biologically-related applications.
Asunto(s)
Elementos de la Serie de los Lantanoides , ADN , Iones , Elementos de la Serie de los Lantanoides/química , Ligandos , Polímeros/químicaRESUMEN
The nucleobase analog 6-thioguanine (6-TG) has emerged as important immunosuppressant, anti-inflammatory, and anticancer drug in the past few decades, but its unique photosensitivity of absorbing strongly ultraviolet UVA light elicits photochemical hazards in many ways. The particularly intriguing yet unresolved question is whether the direct photoreaction of 6-TG can promote DNA-protein cross-links (DPCs) formation, which are large DNA adducts blocking DNA replication and physically impede DNA-related processes. Herein, by real-time observation of radical intermediates using time-resolved UV-vis absorption spectroscopy in conjunction with product analysis by HPLC-MS, we discover that UVA excitation of 6-TG triggers direct covalent cross-linking with tryptophan (TrpH) via an exquisite radical mechanism of electron transfer. The photoexcitation prepares the redox-active triplet 36-TG*, which initiates electron transfer with TrpH, creating TrpHâ¢+ and 6-TGâ¢- in the first step. The deprotonated Trp⢠undergoes radical-recombination with its geminate partner 6-TGâ¢- and eliminates a H2S, leading to the cross-linking product 6-TG-Trp. The photoadduct structures (two chiral isomers and one constitutional isomer) are identified unambiguously, validating further the mechanism. These findings pinpoint the exact amino acid that is vulnerable to photo-cross-linking with 6-TG and establish a mechanistic framework for understanding mutagenic DPCs formation and developing photoprobes based on this new type of photo-cross-linking.
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Tioguanina , Triptófano , ADN , Transporte de Electrón , ElectronesRESUMEN
In this work, we constructed a target-triggered and controlled-release plasmon-enhanced fluorescent AIE probe to realize the purpose of conformational monitoring of insulin fibrillation. We synthesized a novel water-soluble anthracene derivative, 4,4',4'',4'''-(anthracene-9,10-diylbis(ethene-2,1,1-triyl))tetrakis(N,N,N-trimethylbenzenaminium) iodide (BDVAI), with AIE properties, high biocompatibility and good self-assembly effect. Gold nanocages (AuNCs) were selected as the substrate for PEF, and the inner space of hollow AuNCs was filled with BDVAI. Thiol-modified DNA chains were bonded to the surface of AuNCs by Au-S bonds, and an insulin aptamer was combined with the sulfhydryl chain to seal the AuNCs. This PEF-AIE sensor produces different fluorescence signals when interacting with native insulin and fibrillar insulin; thus, monitoring conformational changes in insulin can be realized by detecting fluorescence intensity changes during insulin fibrillation. Based on this design, this system realized sensitive detection of fibrillar insulin with a detection limit of 23.6 pM. This AIE molecular-based PEF fluorescence enhancement system improves the optical properties of fluorescent substances, which is of great significance in improving the detection sensitivity of amyloid fibrils conformational changes and providing a reliable basis for further understanding the pathogenesis of amyloidosis.
Asunto(s)
Técnicas Biosensibles , Colorantes Fluorescentes/química , Insulina/análisis , Colorantes Fluorescentes/síntesis química , Insulina/metabolismo , Conformación MolecularRESUMEN
Bioimaging probes for monitoring intracellular reactive oxygen species have important implications for cell biology research. Herein, we developed peptide-capped silver/gold nanoclusters (peptide@Ag/Au NCs) for lysosome-targeted imaging of hypochlorite (ClO-). The peptide@Ag/Au NCs were synthesized via a one-pot method using peptide as both a template and a reducing agent. The fluorescence intensity and absolute quantum yield of peptide@Ag/Au NCs were much higher than those of peptide-capped gold nanoclusters and silver nanoclusters. In the presence of ClO-, the fluorescence of peptide@Ag/Au NCs was quenched, accompanied by a redshift due to ClO--induced oxidation of the peptide ligand and decreased Ag content in Ag/Au NCs. The relative fluorescence intensity F0/F had favourable linearity for ClO- concentrations in the range 0.1-100 µmol/L (R2 = 0.9954), with a detection limit (LOD) of 80 nmol/L. The lysosome-targeted peptide@Ag/Au NCs were applied to detect ClO- in lysosomes in living cells via fluorescence imaging.
Asunto(s)
Fluorescencia , Ácido Hipocloroso/análisis , Lisosomas/química , Nanopartículas del Metal/química , Imagen Óptica , Péptidos/química , Oro/química , Células Hep G2 , Humanos , Microscopía Fluorescente , Plata/química , Células Tumorales CultivadasRESUMEN
Because of its unique electronic band structure, molybdenum disulfide (MoS2) has been regarded as a star semiconducting material. However, direct growth of continuous and high-quality MoS2 films on SiO2/Si substrates is still very challenging. Here, we report a facile chemical vapor deposition (CVD) method based on synergistic modulation of precursor and Na2SO4 catalysis, realizing the centimeter scale growth of a continuous MoS2 film on SiO2/Si substrates. The as-grown MoS2 film had an excellent spatial homogeneity and crystal quality, with an edge length of the composite domain as large as 632 µm. Both experimental and theoretical results proved that Na tended to bond with SiO2 substrates rather than to interfere with as-grown MoS2. Thus, they showed decent and uniform electrical performance, with electron mobilities as high as 5.9 cm2 V-1 s-1. We believe our method will pave a new way for MoS2 toward real application in modern electronics.
RESUMEN
Human telomeric RNA (TERRA) containing thousands of G-rich repeats has the propensity to form parallel-stranded G-quadruplexes. The emerging crucial roles of TERRA G-quadruplexes in RNA biology fuel increasing attention for studying anticancer ligand binding with such structures, which, however, remains scarce. Here we utilized multiple steady-state and time-resolved spectroscopy analyses in conjunction with NMR methods and investigated thoroughly the binding behavior of TMPyP4 to a TERRA G-quadruplex dimer formed by the 10-nucleotide sequence r(GGGUUAGGGU). It is clearly identified that TMPyP4 intercalates into the 5'-5' stacking interface of two G-quadruplex blocks with a binding stoichiometry of 1:1 and binding constant of 1.92 × 106 M-1. This is consistent with the unique TERRA structural features of the enlarged π-π stacking plane of the A·(G·G·G·G)·A hexad at 5'-ends of each G-quadruplex block. The preferential binding of π-ligand porphyrin to the 5'-5' stacking interface of the native TERRA G-quadruplex dimer is first ascertained by the combination of dynamics and structural characterization.
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G-Cuádruplex , Sustancias Intercalantes/química , Porfirinas/química , ARN/química , Telómero/química , Secuencia de Bases , Dimerización , Humanos , Cinética , Ligandos , Modelos Moleculares , Estructura Molecular , Relación Estructura-ActividadRESUMEN
The Slit-Robo GTPase-activating proteins (srGAPs) are critical for neuronal migration through inactivation of Rho GTPases Cdc42, Rac1, and RhoA. Here we report that srGAP2 physically interacts with protein arginine methyltransferase 5 (PRMT5). srGAP2 localizes to the cytoplasm and plasma membrane protrusion. srGAP2 knockdown reduces cell adhesion spreading and increases cell migration, but has no effect on cell proliferation. PRMT5 binds to the N terminus of srGAP2 (225-538 aa) and methylates its C-terminal arginine residue Arg-927. The methylation mutant srGAP2-R927A fails to rescue the cell spreading rate, is unable to localize to the plasma membrane leading edge, and perturbs srGAP2 homodimer formation mediated by the F-BAR domain. These results suggest that srGAP2 arginine methylation plays important roles in cell spreading and cell migration through influencing membrane protrusion.
Asunto(s)
Movimiento Celular/fisiología , Proteínas Activadoras de GTPasa/metabolismo , Multimerización de Proteína/fisiología , Sustitución de Aminoácidos , Animales , Arginina/genética , Arginina/metabolismo , Células CHO , Membrana Celular/genética , Membrana Celular/metabolismo , Proliferación Celular , Cricetinae , Cricetulus , Citoplasma/genética , Citoplasma/metabolismo , Proteínas Activadoras de GTPasa/genética , Células HEK293 , Células HeLa , Humanos , Metilación , Mutación Missense , Proteína Metiltransferasas/genética , Proteína Metiltransferasas/metabolismo , Proteína-Arginina N-Metiltransferasas , Proteína de Unión al GTP cdc42/genética , Proteína de Unión al GTP cdc42/metabolismo , Proteína de Unión al GTP rac1/genética , Proteína de Unión al GTP rac1/metabolismo , Proteína de Unión al GTP rhoA/genética , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
Maintenance of the Golgi apparatus (GA) structure and function depends on Golgi matrix proteins. The posttranslational modification of Golgi proteins such as phosphorylation of members of the golgin and GRASP families is important for determining Golgi architecture. Some Golgi proteins including golgin-84 are also known to be methylated, but the function of golgin methylation remains unclear. Here, we show that the protein arginine methyltransferase 5 (PRMT5) localizes to the GA and forms complexes with several components involved in GA ribbon formation and vesicle tethering. PRMT5 interacts with the golgin GM130, and depletion of PRMT5 causes defects in Golgi ribbon formation. Furthermore, PRMT5 methylates N-terminal arginines in GM130, and such arginine methylation appears critical for GA ribbon formation. Our findings reveal a molecular mechanism by which PRMT5-dependent arginine methylation of GM130 controls the maintenance of GA architecture.