ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 11
·2487-2494
·
IF 3.637
Xue Ning,Wu Shujie,Li Zongbing,Miao Xiangmin
A versatile nanocomposite was simply prepared based upon the electrostatic adsorption of positively charged gold nanoparticles with negatively charged graphene oxide ( nano-gold@GO), and utilized as a novel fluorescence quenching platform for ultrasensitive detection of adenosine triphosphate (ATP). In the designed system, DNA-stabilized Ag nanoclusters (DNA/AgNCs) were used as fluorescent probes, DNA duplex was formed in the presence of ATP, and they can electrostatically adsorb onto the surface of nano-gold@GO to quench the fluorescence signal. Upon the addition of exonuclease III (Exo III), the DNA duplex would be hydrolyzed into DNA fragments and resulted in the recovery of the fluorescence signals due to the diffusion of AgNCs away from nano-gold@GO. Based on these, sensitive detection of ATP was realized with a detection range of 5.0 pM–20 nM. Notably, a good recovery in the range of 94–104% was obtained when detecting ATP in human serum samples, indicating a promising application value in early disease diagnosis.
A functional positively charged nano-gold@graphene oxide was fabricated and utilized as an enhanced fluorescence quenching platform for the detection of ATP, coupled with exonuclease III-assisted signal amplification.
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FEBS Journal
·
2018
·Vol. 285
·期 12
·2292-2305
·
IF 4.53
Jin-Hua Guo,Ren-Wang Jiang,Jacob Lauwring Andersen,Mikael Esmann,Natalya U. Fedosova
The information obtained from crystallized complexes of the Na + ,K + -ATPase with cardiotonic steroids (CTS) is not sufficient to explain differences in the inhibitory properties of CTS such as stereoselectivity of CTS binding or effect of glycosylation on the preference to enzyme isoforms. The uncertainty is related to the spatial organization of the hydrophilic cavity at the entrance of the CTS-binding site. Therefore, there is a need to supplement the crystallographic description with data obtained in aqueous solution, where molecules have significant degree of flexibility. This work addresses the applicability of the electron paramagnetic resonance (EPR) method for the purpose. We have designed and synthesized spin-labeled compounds based on the cinobufagin steroid core. The length of the spacer arms between the steroid core and the nitroxide group determines the position of the reporting group (N-O) confined to the binding site. High affinity to Na + ,K + -ATPase is inferred from their ability to inhibit enzymatic activity. The differences between the EPR spectra in the absence and presence of high ouabain concentrations identify the signature peaks originating from the fraction of the spin labels bound within the ouabain site. The degree of perturbations of the EPR spectra depends on the length of the spacer arm. Docking of the compounds into the CTS site suggests which elements of the protein structure might be responsible for interference with the spin label (e.g., steric clashes or immobilization). Thus, the method is suitable for gathering information on the cavity leading to the CTS-binding site in Na + ,K + -ATPase in all conformations with high affinity to CTS .
研究领域:
生物化学
·
药理学
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Jinye Huang,Xuan Xuan,Dongmei Xu,Yuezhong Wen
The regulation solutions and mechanisms of reducing pesticide phytotoxicity to nontarget plants are not well-defined and detailed. Here, we have proposed a new detoxification strategy to control the toxic effects of herbicide imazethapyr (IM) induced in wheat seedlings from the perspective of the plasma membrane (PM) H+-ATPase. We found that the changes in PM H+-ATPase activity have a regulatory effect on the phytotoxic effects induced by IM in plants. Treatment with PM H+-ATPase activators restored the reduced auxin content and photosynthetic efficiency caused by IM, thereby promoting plant growth. Application of a PM H+-ATPase inhibitor further reduced phosphorus content and significantly increased 2,4-dihydroxy-7-methoxy-2H,1,4-benzoxazin-3(4H)one (DIMBOA) and jasmonic acid levels. These effects indicate that auxin and DIMBOA may regulate plant growth trends and detoxification effects mediated by PM H+-ATPase. This work opens a new strategy for regulating herbicide toxicity to nontarget plants from the PM H+-ATPase.
研究领域:
植物生理学
·
植物生物化学
·
农药科学
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Journal of Cleaner Production
·
2024
·
IF 9.7
Linya Yi,Shuyan Chen,Yuancai Chen
Bio-remediation are low-cost and environmentally friendly, but microbial enzymes tend to lose their activity when treated antibiotic contaminants due to its high toxicity. In this study, Bacillus megaterium Y-4 was demonstrated to in situ synthesize Pd 0 @cells, and detoxify oxytetracycline (OTC) through enhancing electron transport chain via Pd 0 -dependent F O F 1 -ATPase (proton pumps). Toxicity analyses showed that increasing Pd 0 loading significantly reduced acute and chronic bio-toxicity as well as ecological risk of OTC effluent, as evidenced by the promoting cellular response to oxidative stress with increasing enzymes GSH-Px and IDH activities (4.24 and 1.20-fold). Electrochemical (I-t, LSV, DPV and EDC) showed that the incorporated bio-Pd 0 mainly promoted FDH/Hase-based short chain (gradually took precedence over CoQ-based long chain), diverting OTC extracellular degradation to markedly detoxify OTC to cell. Inhibition (DCCD and TBA) assays and structural equation modelling indicated that bio-Pd 0 upregulated ATP and NADH synthesis. It also acted as transmembrane F O F 1 -ATPase proton pump, which enabled the backward of protons out of the restriction of PMF , eventually maximizing the energy utilization. This study sheds new insights into the detoxification of antibiotics by bio-nanoparticles through proton transmembrane transfer/redistribution.
研究领域:
微生物学
·
环境科学
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ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Fei Peng,Xiangnan Ai,Jing Sun,Xichuan Ge,Meiqi Li,Peng Xi,Baoxiang Gao
Mitochondrial cristae, invaginations of the inner mitochondrial membrane (IMM) into the matrix, are the main site for the generation of ATP via oxidative phosphorylation, and mitochondrial membrane potential (MMP). Synchronous study of the dynamic relationship between cristae and MMP is very important for further understanding of mitochondrial function. Due to the lack of suitable IMM probes and imaging techniques, the dynamic relationship between MMP and cristae structure alterations remains poorly understood. We designed a pair of FRET-based molecular probes, with the donor (OR-LA) being rhodamine modified with mitochondrial coenzyme lipoic acid and the acceptor (SiR-BA) being silicon-rhodamine modified with a butyl chain, for simultaneous dynamic monitoring of mitochondrial cristae structure and MMP. The FRET process of the molecular pair in mitochondria is regulated by MMP, enabling more precise visualization of MMP through fluorescence intensity ratio and fluorescence lifetime. By combining FRET with FLIM super-resolution imaging technology, we achieved simultaneous dynamic monitoring of mitochondrial cristae structure and MMP, revealing that during the decline of MMP, there is a progression involving cristae dilation, fragmentation, mitochondrial vacuolization, and eventual rupture. Significantly, we successfully observed that the rapid decrease in MMP at the site of mitochondrial membrane rupture may be a critical factor in mitochondrial fragmentation. These data collectively reveal the dynamic relationship between cristae structural alterations and MMP decline, laying a foundation for further investigation into cellular energy regulation mechanisms and therapeutic strategies for mitochondria-related diseases.
研究领域:
细胞生物学
·
生物物理学
·
医学影像学
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ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Mengke Wang,Zihan Lin,Qing Liu,Shan Jiang,Hua Liu,Xingguang Su
A novel fluorescent biosensor for protein kinase activity (PKA) detection was designed by applying double-strands DNA-hosted copper nanoclusters (dsDNA-CuNCs) and graphene oxide (GO). One DNA strand of the dsDNA consisted of two domains, one domain can hybridize with another complementary DNA strand to stabilize the fluorescent CuNCs and another domain was adenosine 5′-triphosphate (ATP) aptamer. ATP aptamer of the dsDNA-CuNCs would be spontaneously absorbed onto the GO surface through π-π stacking interactions. Thus GO can efficiently quench the fluorescence (FL) of dsDNA-CuNCs through fluorescence resonance energy transfer (FRET). In the present of ATP, ATP specifically combined with ATP aptamer to form ATP-ATP aptamer binding complexes, which had much less affinity to GO, resulting in the fluorescence recovery of the system. Nevertheless, in the presence of PKA, ATP could be translated into ADP and ADP could not combine with ATP aptamer resulting in the fluorescence quenching of dsDNA-CuNCs again. According to the change of the fluorescence signal, PKA activity could be successfully monitored in the range of 0.1–5.0 U mL −1 with a detection limit (LOD) of 0.039 U mL −1 . Besides, the inhibitory effect of H-89 on PKA activity was studied. The sensor was performed for PKA activity detection in cell lysates with satisfactory results.
研究领域:
生物化学
·
分子生物学
·
分析化学
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ACS Nano
·
2019
·
IF 13.903
Yi Du,Pai Peng,Tao Li
DNA logic nanodevices that in situ operate within living cells have attracted increasing interest and shown great promise for gene regulation and target recognition. A challenge remains how to control their activation inside specific cellular compartments. Toward this goal, here we report a lysosome-recognizing framework nucleic acid (FNA) nanodevice using an i-motif and an ATP-binding aptamer (ABA) incorporated into a DNA triangular prism (DTP) as the logic-controlling units. Once entering the lysosomal compartments, the FNA device responds to lysosomal pH and ATP via the folding of i-motif and ABA, which triggers a structural change of FNA and the release of a reporter structure for subcellular imaging. With endogenous proton and ATP as two inputs, an AND logic gate is built and in situ operated within living lysosomes by pH and ATP modulation with external drug stimuli. Given the abnormal levels of pH and ATP within some cancer cells or dysfunctional lysosomal cells, in this context our designed FNA logic device may find extended applications in controllable drug release and disease treatment.
研究领域:
分子生物学
·
纳米医学
·
细胞生物学
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FEBS LETTERS
·
2015
·Vol. 590
·期 1
·22-33
·
IF 3.169
Xue Fu,Xingjie Gao,Lin Ge,Xiaoteng Cui,Chao Su,Wendong Yang,Xiaoming Sun,Wei Zhang,Zhi Yao,Xi Yang,Jie Yang
Malonate, a classic inhibitor of respiratory electron transport chain, induces mitochondrial stress. Stress granules (SGs) are a kind of dynamic foci structure during stress. The study on the connection of mitochondrial stress and SG assembly is still limited. Here, we demonstrated that malonate treatment leads to SG formation and translation inhibition, apart from mitochondrial stress, including enhanced ROS formation, reduced mitochondrial Δψ m and ATP level. The phosphorylation levels of eIF2α and 4EBP1 protein were affected upon mitochondrial dysfunction. However, knockdown of 4EBP1 affected SG formation, rather than eIF2α. In addition, an increase of ATP level under mitochondrial stress enhanced malonate-induced SG aggregation. Overall, malonate stimulation triggers mitochondrial stress and induces the assembly of non-canonical cellular SGs via 4EBP1 pathway.
研究领域:
细胞生物学
·
分子生物学
·
生物化学
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METABOLIC ENGINEERING
·
2019
·
IF 7.808
Ting Shi,Shan Liu,Yi-Heng P. Job Zhang
Carbon dioxide (CO 2 ) is an appealing carbon feedstock for the sustainable production of biocommodities. Here we designed three in vitro artificial enzymatic pathways featuring the ATP-excess, ATP-deficit, and ATP-balanced pathways for the biotransformation of starch and CO 2 to malate. This ATP-balanced pathway without exogenous ATP donors can auto-regulate its carbon fluxes from glyceraldehyde 3-phosphate to 3-phosphoglycerate via either the ATP-generating pathway (a part of glycolysis) or no-ATP-generating pathway from a hyperthermophilic archaeon Thermococcus kodakarensis . The ATP-balanced pathway enabled to produce up to 52.4 mM malate with 95.3% of the theoretical yield, that is, 2 mol of malate synthesized from 1 mol of glucose of starch and 2 mol of CO 2 . This pathway also enabled to produce high-yield malate regardless of ATP/ADP ratios. Anaerobic reaction conditions and/or the addition of a reducing agent dithiothreitol were of importance for creating an anoxic environment for biocatalysis of enzyme cocktails and for mitigating the deactivation of enzymes and degradation of intermediates. This new pathway could provide a green route for direct conversion of CO2 to many building blocks, a promising alternative of petrochemical-based production of biocommodities.
研究领域:
生物化学
·
合成生物学
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Scientific Reports
·
2017
·Vol. 7
·期 1
·1-8
·
IF 4.259
Feng Ruizhi,Shi Weining,Wang Dejia,Wen Jia,Li Hongjuan,Sun Shiguo,Xu Yongqian
Optical activity of hierarchical supramolecular assemblies based on organic dyes would create multiple functional architectures. In this work, three kinds of silica nanoparticles with or without functional groups were synthesized. For the first time, silica nanoparticles can induce positively charged squaraine (SQ) to aggregate to form supramolecular assemblies. Adenosine-5′-triphosphate (ATP) as building blocks was absorbed on the surface of silica nanoparticles through metal-anion coordination and electrostatic interactions, in which the aggregates of SQ was transferred to monomer. The thickness being composed of ATP and SQ on the outside of nanoparticles is about 5 nm. These supramolecular assemblies showed selective turn-on fluorescence response to ATP in near infrared (NIR) region over other ions through metal-anion coordination and electrostatic interactions. These functional silica nanoparticles possessing many advantages provide proof-of-principle “seed crystals” for construction of supramolecular assemblies and platforms for sensing with facile performance.
研究领域:
材料科学
·
生物化学
·
分析化学
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ADVANCED FUNCTIONAL MATERIALS
·
2025
·
IF 19
Zhengtao Xu,Chen Wang,Zhuangzhuang Zhao,Yuxin Zhang,Yao Ying,Juan Li,Liang Qiao,Jingwu Zheng,Shenglei Che,Jing Yu
Theranostic probes that integrate diagnosis and treatment together are considered as effective tools for individual therapy. However, the separation of detection and treatment components limits precise assessment of therapeutic efficacy. Herein, a novel self-monitoring probe that sensitively responds to adenosine triphosphate (ATP) was developed by ultrafine iron oxide nanowires (UIONWs) through an oleyl amine-assisted way. The UIONWs, with a diameter of 1.5 nm and length of 75 nm, are surface-coordinated with oleyl amine. UIONWs can be decomposed in ATP-containing solution and expose the shielded iron ions. Consequently, it can boost the peroxidase (POD) activity and activate the T 1 -magnetic resonance imaging (MRI) signal by ATP. In addition, UIONWs can be rapidly engulfed into cells due to the high aspect ratio. The improved ATP and hydrogen peroxide (H 2 O 2 ) within tumor cells leads to higher hydroxyl radicals (OH) generation for effective tumor inhibition.In contrast, normal cells exhibit lower ATP and H 2 O 2 levels, reducing cytotoxicity and ensuring safety. Since both the illumination of T 1 -MRI and OH generation are ATP-controlled, this work raised a potential way to self-monitor tumor therapy with timely feedback on therapeutic effects.
研究领域:
生物医学工程
·
纳米医学
·
影像医学
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 11
·1-8
·
IF 5.479
Guo Ying,Zhang Juncai,Zhang Weihong,Hu Daodao
Brightly fluorescent carbon quantum dots coated with polyethylenimine (PEI-CDs) were prepared using malic acid and PEI as the precursors. The PEI-CDs have a high quantum yield (41%) and green emission (peaking at 502 nm under 430 nm excitation), both of which are not affected by high ionic strength. The PEI-CDs have a positive charge at physiological pH values and can electrostatically bind aptamers with their negative charge. This is shown for aptamers binding thrombin or ATP. Binding of aptamers results in quenching of fluorescence. If thrombin or ATP are introduced, the respective aptamer will bind them, and the complex is then released from the PEI-CDs. Fluorescence increases in proportion to the analyte concentration. Under optimized conditions, thrombin and ATP can be sensitively and selectively detected by fluorometry with lower detection limits of 1.2 and 13 nM, respectively. The assay was successfully applied to the determination of thrombin and of ATP in spiked serum samples.
Green fluorescent carbon quantum dots were functionalized with polyethyleneimine. They were applied to aptamer-based determination of thrombin and ATP. The PEI-functionalized carbon quantum dots (PEI-CDs) have bright green fluorescence are were synthesized by one-step hydrothermal treatment of malic acid and PEI. Employing the PEI-CDs, a fluorometric aptamer-based assay was developed for the determination of thrombin and ATP.
研究领域:
分析化学
·
生物化学
·
纳米技术
·
分子生物学
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