BIOSENSORS & BIOELECTRONICS
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2013
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IF 5.437
Bingqian Liu,Bing Zhang,Guonan Chen,Huanghao Yang,Dianping Tang
A new flow-through electrochemical aptasensor was designed for ultrasensitive monitoring of adenosine triphosphate (ATP) by coupling microvalve-programmable capillary column with CdS-functionalized DNA concatamer for signal amplification . Initially, a layer of primary DNA-conjugated polyacrylamide hydrogel was covalently linked onto the internal surface of capillary column, and then an automated sequenctial injection format with a syringe pump was employed for development of the programmable capillary-based aptasensor. In the presence of target DNA aptamer, the immobilized primary DNA hybridized with partial bases of the aptamer. The excess aptamer fregment could trigger the formation of DNA concatamer between auxiliary DNA 1 and CdS-labeled auxiliary DNA 2 . Upon target ATP introduction, a specific ATP-aptamer reaction was excuated, thereby resulting in the release of CdS-functionalized DNA concatamer from the capillary. Subsenquent anodic stripping voltammetric detection of cadmium released under acidic conditions from the released CdS nanoparticles could be conducted in a homemade detection cell. Under optimal conditions, the dynamic concentration range spanned from 0.1 pM to 10 nM ATP with a detection limit of 0.06 pM ATP. The electrochemical aptasensor showed good reproducibility, selectivity, and stability. In addition, the methodology was evaluated for the analysis of ATP spiked serum samples, and the recoveries was 81–140%.
研究领域:
生物化学
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分析化学
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分子生物学
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DOI
PubMed
高影响因子
RSC Advances
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2024
·Vol. 14
·期 8
·5594-5599
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IF 3.9
Yuxia Li,Zeting Meng,Yating Liu,Baozhu Zhang
A turn-on fluorescence nanoprobe was constructed for the determination of adenosine 5′-triphosphate (ATP) based on DNA-templated silver nanoclusters (DNA-AgNCs). The significant enhancement fluorescence intensity of DNA-AgNCs in the presence of ATP is due to the high special binding affinity between ATP and the aptamer, resulting in the environment of DNA-AgNCs with darkish fluorescence lying at one terminus of DNA slightly altering owing to the change of ATP aptamer conformation. A good linear range runs from 9 to 24 mM with a satisfactory detection limit of 3 μM. Furthermore, the proposed nanoprobe exhibited good performance for ATP detection in diluted fetal bovine serum.
研究领域:
分析化学
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纳米技术
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生物化学
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DOI
PubMed
高影响因子
Polymers
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2019
·Vol. 11
·期 7
·1139
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IF 3.164
Lihua Liu,Linlin Zhao,Dandan Cheng,Xinyi Yao,Yan Lu
A boronic acid groups-bearing polythiophene derivate (L) was designed and synthesized for highly sensitive fluorescence detection of ATP based on a multisite-binding coupled with analyte-induced aggregation strategy.Lhas a polythiophene backbone as fluorophores and two functional side groups, i.e., quaternary ammonium group and boronic acid group, as multibinding sites for ATP. When various structural analogues such as ADP, AMP, and various inorganic phosphates were added into the aqueous solution ofL, only ATP caused a remarkable fluorescence quenching of about 60-fold accompanied by obvious color changes of solution from yellow to purple. The detection limit is estimated to be 2 nM based on 3σ/slope. With the advantage of good water solubility, low toxicity, and highly selective response to ATP,Lwas successfully utilized as a probe to real-time assay activity of adenylate kinase (ADK) and map fluorescent imaging of ATP in living cells.Keywords:ATP detection;fluorescent probe;polythiophene;multisite binding;analyte-induced aggregation
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2023
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IF 4.4
Jabrane Jouha,Fengli Li,Hai Xiong
Due to their distinctive physical, chemical, electrical, and optical properties as well as their prospective uses, 2D covalent organic framework (COF) have attracted much attention. Herein, TaTPA-COF was effectively synthesized from the condensation of TTA and TFPA using a facile solvothermal method and characterized by SEM image, FT-IR spectra, and PXRD pattern. The generated bulk TaTPA-COF materials combined with DNA aptamers are utilized as the acceptor (quencher) for the highly sensitive and selective detection of adenosine 5′-triphosphate (ATP) and thrombin, with a novel fluorescence biosensing platform and a proof-of-concept application.
研究领域:
生物化学
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纳米医学
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分析化学
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DOI
PubMed
高影响因子
Small
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2010
·Vol. 7
·期 1
·87-94
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IF 6.171
Yi-Lun Ying,Hai-Yan Wang,Todd C. Sutherland,Yi-Tao Long
An aptamer is a specific oligonucleotide sequence that spontaneously forms a secondary structure capable of selectively binding an analyte. An aptamer’s conformation is the key to specific binding of a target molecule, even in the case of very closely related targets. Nanopores are a sensitive tool for the single-molecule analysis of DNA, peptides, and proteins transporting through the pore. Herein, a single α -hemolysin natural nanopore is utilized to sense the conformational changes of an adenosine 5’-triphosphate (ATP)-binding aptamer (ABA). The known DNA sequence of the ABA is used as a model to develop real-time monitoring of molecular conformational changes that occur by binding targets. The native, folded ABA structure has a nanopore unfolding time of 4.17 ms, compared with 0.29 ms for the ABA:ATP complex. A complementary 14-mer strand, which binds the ABA sequence in the key nucleic acids responsible for folding, forms linear duplex DNA, resulting in a nanopore transit time of 0.50 ms and a higher capture probability than that of the folded ABA oligomer. Competition assays between the ABA:ATP and ABA:reporter complexes are carried out, and the results suggest that the ABA:ATP complex is formed preferentially. The nanopore allows for the detection of an ABA in its folded, ATP-bound, and linear conformations.
研究领域:
分子生物学
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生物传感器
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核酸化学
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单分子检测
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转化医学
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DOI
高影响因子
SKIN RESEARCH AND TECHNOLOGY
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2023
·Vol. 29
·期 3
·e13303
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IF 2.2
Chunxia Gan,Titi Liu,Xiaorong Jia,Xueqin Huang,Xiangdong Qin,Xuanjun Wang,Jun Sheng,Huanhuan Xu
Background Skin photoaging is the damage caused by excessive exposure to ultraviolet (UV) irradiation. We investigated the effect of adenosine triphosphate (ATP) supplementation on UVB-induced photoaging in HaCaT cells and its potential molecular mechanism. Materials and methods The toxicity of ATP on HaCaT cells was examined by the MTT assay. The effects of ATP supplementation on the viability and apoptosis of HaCaT cells were determined by crystal-violet staining and flow cytometry, respectively. Cellular and mitochondrial ROS were stained using fluorescent dyes. Expression of Bax, B-cell lymphoma (Bcl)-2, sirtuin (SIRT)3, and superoxide dismutase (SOD)2 was measured via western blotting. Results ATP (1, 2 mM) exerted no toxic effect on the normal growth of HaCaT cells. UVB irradiation caused the apoptosis of HaCaT cells, and ATP supplementation inhibited the apoptosis induced by UVB significantly, as verified by expression of Bax and Bcl-2. UVB exposure resulted in accumulation of cellular and mitochondrial reactive oxygen species (ROS), but ATP supplementation suppressed these increases. Expression of SIRT3 and SOD2 was decreased upon exposure to UVB irradiation but, under ATP supplementation, expression of SIRT3 and SOD2 was reversed, which was consistent with the reduction in ROS level observed in ATP-treated HaCaT cells after exposure to UVB irradiation. Conclusions ATP supplementation can suppress UVB irradiation-induced photoaging in HaCaT cells via upregulation of expression of SIRT3 and SOD2.
研究领域:
细胞生物学
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分子生物学
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皮肤科
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DOI
PubMed
高影响因子
ANALYST
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2018
·Vol. 143
·期 16
·3821-3828
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IF 3.864
Chuanxia Chen,Qun Yuan,Pengjuan Ni,Yuanyuan Jiang,Zhenlu Zhao,Yizhong Lu
Alkaline phosphatase (ALP) is a significant biomarker for diagnostics. Simple, selective and sensitive detection of ALP activity is thus of critical importance. In this study, an artful fluorescence assay for ALP is proposed based on adenosine triphosphate (ATP) hydrolysis-triggered disassociation and fluorescence quenching of cerium coordination polymer nanoparticles (CPNs). ATP, a recognized natural substrate of phosphatase, can serve as a superb “antenna” to sensitize the luminescence of Ce3+ with the aid of tris(hydroxymethyl) aminomethane (Tris), forming Ce3+–ATP–Tris CPNs. In the presence of ALP, ATP will be catalytically converted into adenosine and inorganic orthophosphate, however neither of them can sensitize Ce3+ in alkaline media. As a result, the obtained CPNs are disassociated, inducing the quenching of the fluorescence. On this basis, a straightforward fluorescence assay for ALP activity is rationally developed. The fluorescence quenching efficiency shows a linear relationship for ALP within the activity range from 0.1 to 10 mU mL−1 with a detection limit of 0.09 mU mL−1 under the optimal experimental conditions. Moreover, this facile yet effective fluorescence method featured simplicity, cost-effectiveness, high sensitivity and high selectivity and can be successfully utilized for the quantitative detection of ALP in human serum samples.
研究领域:
生物化学
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医学诊断
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纳米技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
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2017
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IF 4.162
Yujie Wang,Xinxin Shang,Jia Liu,Yingshu Guo
Here, we have developed a facile fluorometric system for the detection of adenosine triphosphate (ATP) by a rolling circle amplification (RCA) based on proximity ligation mediated amplification, and simultaneously achieved the release of the anticancer drug doxorubicin (DOX) through the mesoporous silicon system. Once the ATP molecule is present, the linker DNA will be released from the graphene oxide (GO) surface and hybridized to the template DNA of the GO surface joining with ligation enzyme. RCA reaction is followed by the addition of the phi29 DNA polymerase. The product of RCA reaction contains a base fragment complementary to the signal DNA, allowing the fluorescent oligonucleotide probe to be released from the GO surface and fluorescence is recovered. The strong fluorescence signal realized the sensitive detection of ATP. Gate DNA were modified to the surface of the mesoporous silica (MSN) by electrostatic attraction to encapsulate DOX. After the above-mentioned RCA process, its result that long DNA chain containing a base fragment complementary to gate DNA, would be hybridized to the gate DNA strand on the surface of MSN, which opened the MSN hole and released the drug DOX into cell for HeLa cell therapy. And the specificity to folate receptor overexpressed on cell surface was satisfactory which would be beneficial for cancer therapy.
研究领域:
生物化学
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分子生物学
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纳米医学
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DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2025
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IF 3.3
Zhao Bingshan,Micklefield Jason,Wang Yonghua,Wang Fanghua
N-trans -feruloyltyramine (FLA) is one kind of phenylpropanoid compound found in various plants. Numerous studies have confirmed that it exhibits a wide range of physiological functions, such as antioxidant, ɑ-glucosidase inhibition, and anti-inflammatory activity. However, the low content of FLA in plants greatly limits its potential use in food and pharmaceutical industries. It is, therefore, very important to establish an effective synthesis of FLA. In this study, a green and efficient method to synthesize FLA was sought using an amide bond synthetase (ABS) biocatalyst. Ten kinds of ABS enzymes, including AlCfaL from Azospirillum lipoferum , were screened as the potential biocatalysts for the production of FLA. To obtain optimum reaction conditions, the effects of various parameters on conversion of FLA were firstly evaluated. Under the optimum conditions using 1 mM N-trans -ferulic acid, 50 mM tyramine (substrate ratio of 1:50), 10 mM MgCl₂, 8 mM ATP, and 35 µM AlCfaL enzyme at 30 °C with a shaking speed of 500 r/min for 48 h, maximum conversion rate of 74% was reached. Given that the amidation reaction is mediated by relative expensive ATP, we further optimized reaction systems to incorporate an ATP recycling system consisting of a polyphosphate kinase enzyme (CHU) and an inexpensive polyphosphate (PolyP) as the phosphate donor. Response surface methodology (RSM) based on five-level, five-variable central composite design (CCD) was used to evaluate the optimal parameters for the production of FLA. The effects of AMP, PolyP, AlCfaL, CHU concentrations, and reaction time on the conversion rate of FLA were analyzed. The optimum conditions derived via RSM were 7.12 mM AMP, 5.96 mg/mL PolyP, 39.72 μM AlCfaL, 27.68 μM CHU, and a reaction time of 36 h. Validation experiments conducted under these optimized conditions yielded an actual conversion rate of 63.5%, which compared well to the maximum predicted value of 64.2%.
研究领域:
生物化学
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酶工程
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食品科学
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药学
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DOI
PubMed
高影响因子
Biotechnology for Biofuels and Bioproducts
·
2023
·Vol. 16
·期 1
·1-13
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IF 0
Gao Hui,Li Mengxuan,Wang Qing,Liu Tingting,Zhang Xian,Yang Taowei,Xu Meijuan,Rao Zhiming
ATP, an important cofactor, is involved in many biocatalytic reactions that require energy. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We determined the catalytic properties of PPK from different sources and found that PPK from Cytophaga hutchinsonii ( Ch PPK) had the best catalytic activity for the substrates ADP and polyP 6 . An extracellular–intracellular dual system was constructed to high-throughput screen for better catalytic activity of Ch PPK mutants. Finally, the specific activity of Ch PPK D82N-K103E mutant was increased by 4.3 times. Therefore, we focused on the production of L-theanine catalyzed by GMAS as a model of ATP regeneration. Supplying 150 mM ATP, GMAS enzyme could produce 16.8 ± 1.3 g/L L-theanine from 100 mM glutamate. When 5 mM ATP and 5 U/mL Ch PPK D82N-K103E were added, the yield of L-theanine was 16.6 ± 0.79 g/L with the conversion rate of 95.6 ± 4.5% at 4 h. Subsequently, this system was scaled up to 200 mM and 400 mM glutamate, resulting in the yields of L-theanine for 32.3 ± 1.6 g/L and 62.7 ± 1.1 g/L, with the conversion rate of 92.8 ± 4.6% and 90.1 ± 1.6%, respectively. In addition, we also constructed an efficient ATP regeneration system from glutamate to glutamine, and 13.8 ± 0.2 g/L glutamine was obtained with the conversion rate of 94.4 ± 1.4% in 4 h after adding 6 U/ mL GS enzyme and 5 U/ mL Ch PPK D82N-K103E , which further laid the foundation from glutamine to L-theanine catalyzed by GGT enzyme. This proved that giving the reaction an efficient ATP supply driven by the mutant enzyme enhanced the conversion rate of substrate to product and maximized the substrate value. This is a positively combination of high yield, high conversion rate and high economic value of enzyme catalysis. The mutant enzyme will further power the ATP-consuming biocatalytic reaction platform sustainably.
研究领域:
生物化学
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酶工程
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代谢工程
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DOI
PubMed
SENSORS AND ACTUATORS B-CHEMICAL
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2024
·
IF 8
Jiarong Lv,Qinchuan Wei,Xuewen Gong,Enming Du,Shijin Zhang
Extracellular adenosine triphosphate (ATP) plays a crucial role in recruiting neutrophils, which then release neutrophil elastase (NE) to address inflammatory stimuli. However, there has been no prior report on the simultaneous measurement of ATP and NE levels under inflammatory conditions. Herein, we have developed a dual-color molecular fluorescent probe, termed AN, that allows for the simultaneous detection of ATP and NE, providing insights into their interactions during inflammation. AN demonstrates excellent sensitivity and minimal spectral crosstalk, with distinct detection channels for ATP (λ ex = 540 nm, λ em = 583 nm) and NE (λ ex = 450 nm, λ em = 531 nm). It shows high selectivity in sensing ATP and NE within cell cultures, even amidst complex biological environments. Notably, AN effectively distinguishes the progression of inflammatory conditions in vivo . Our findings reveal a direct correlation between ATP and NE levels in inflamed tissues and offer valuable insights for monitoring inflammation and guiding clinical interventions.
研究领域:
生物化学
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分子生物学
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炎症研究
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荧光探针
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DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2024
·
IF 4.3
Ruo-Fei Ma,Qi Zhang,Yue Wang,Zhang-Run Xu
An ultrasensitive strategy for in-situ visual monitoring of ATP in a single living tumor cell during mitochondria-targeted photodynamic therapy (PDT) process with high spatiotemporal resolution was proposed using surface-enhanced Raman scattering (SERS) 3D imaging technique. The nanostructures consisting of Au-Ag 2 S Janus nanoparticles functionalized with both Au nanoparticles linked by a DNA chain and a mitochondrial-targeting peptide (JMDA NPs) were deliberately employed to target mitochondria. The JMDA NPs exhibit excellent SERS activity and remarkable antitumor activity. The quantization of ATP relies on the intensity of the SERS probes bonded to the DNA, which shows a strong correlation with the generated hot spot between the Janus and the Au. Consequently, spatiotemporally controlled monitoring of ATP in the mitochondria of single living cells during the PDT process was achieved. Additionally, the JMDA NPs demonstrated remarkable capability for mitochondria-targeted PDT, providing significant antitumor effects and superior therapeutic safety both in vitro and in vivo . Our work presents an effective JMDA NPs-based SERS imaging strategy for in-situ and real-time 3D visualization of intracellular ATP in living tumor cells during the mitochondria-targeted PDT process, which enables significant information on the time point of PDT treatment and is beneficial to precious PDT applications in tumor therapy.
研究领域:
生物医学工程
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肿瘤学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
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