CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2024
·
IF 1.2
Liyi Ma,Lixuan Mu,Lushan Huang,Guangwei She,Wensheng Shi
An outstanding fluorescent probe for adenosine 5′-triphosphate (ATP) in the millimolar range will shed deep light on the physiological role of ATP. Many fluorescent probes based on rhodamine spirolactam derivatives have been developed for this purpose and claimed to detect millimolar ATP excellently, where ATP or adenosine 5′-triphosphate disodium salt (ATP-2Na) dissolved in a buffer solution is employed to examine their response to ATP. However, in this work, it was found that both millimolar ATP and ATP-2Na could significantly decrease the pH of the common buffer solution even for several units. Considering the dependence of the rhodamine spirolactam fluorescence on pH, a series of rhodamine spirolactam-based probes was synthesized to investigate their fluorescence response to pH and ATP-2Na. The results demonstrated that the fluorescence response of the probes to millimolar ATP-2Na indeed resulted from both ATP-2Na itself and the pH decrease induced by ATP-2Na. Furthermore, it was evidenced that acidic pH was crucial for the probes to produce fluorescence response to millimolar ATP-2Na itself, and the rhodamine spirolactam-based probes would show poor sensitivity to millimolar ATP-2Na at near-natural pH. Accordingly, the previous conclusion that the rhodamine spirolactam-based probes could detect millimolar ATP excellently was almost erroneous, and might mislead the design and practice of intracellular ATP probe in the future. This work notes that attention should be paid to the physiochemical properties of the analyte and the environmental changes produced by the target analyte.
研究领域:
分析化学
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生物化学
·
分子生物学
·
荧光光谱学
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产品关联
DOI
高影响因子
Scientific Reports
·
2017
·
IF 4.259
Xie Xiaoyun,Han Chaofei,Zeng Weiqi,Chen Chen,Lu Lixia,Liu Queping,Peng Cong,Zhao Shuang,Su Juan,Chen Xiang
The F1F0-ATP synthase, an enzyme complex, is mainly located on the mitochondrial inner membrane or sometimes cytomembrane to generate or hydrolyze ATP, play a role in cell proliferation. This study focused on the role of F1F0-ATP synthase in keratinocyte differentiation, and its relationship with intracellular and extracellular ATP (InATP and ExATP). The F1F0-ATP synthase β subunit (ATP5B) expression in various skin tissues and confluence-dependent HaCaT differentiation models was detected. ATP5B expression increased with keratinocyte and HaCaT cell differentiation in normal skin, some epidermis hyper-proliferative diseases, squamous cell carcinoma, and the HaCaT cell differentiation model. The impact of InATP and ExATP content on HaCaT differentiation was reflected by the expression of the differentiation marker involucrin. Inhibition of F1F0-ATP synthase blocked HaCaT cell differentiation, which was associated with a decrease of InATP content, but not with changes of ExATP. Our results revealed that F1F0-ATP synthase expression is associated with the process of keratinocyte differentiation which may possibly be related to InATP synthesis.
研究领域:
细胞生物学
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分子生物学
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皮肤病学
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DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Fangfang Li,Xuan Hu,Fengyi Wang,Baozhan Zheng,Juan Du,Dan Xiao
A simple, rapid, and ultrasensitive fluorescence strategy for adenosine triphosphate (ATP) detection was developed by using a FAM (carboxyfluorescein) labeled DNA (FAM-DNA). In this strategy, highly fluorescent FAM-DNA was used as a probe, and nanoceria (CeO 2 NPs) acted as an efficient quencher. FAM-DNA attached to the surface of nanoceria through the coordination between the phosphate group of DNA and NP surface, which induced complete quenching in the FAM-DNA fluorescence due to a photo induced electron transfer (PET) process. It was found that ATP can readily displace adsorbed DNA from nanoceria surface because of the stronger coordination ability of ATP with nanoceria, and the nanoceria-based competitive binding resulted in over 7-fold fluorescence enhancement. Over a wide range from 0.1 nM to 1.5 μM, a good linear relationship between the fluorescence intensity and the concentration of ATP was obtained and the detection limit was estimated to be as low as 54 pM. This method was successfully used to analyze ATP in a single drop of blood and human urine.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
Bioresources and Bioprocessing
·
2021
·Vol. 8
·期 1
·1-10
·
IF 4.578
Sun Chuanqi,Li Zonglin,Ning Xiao,Xu Wentian,Li Zhimin
Adenosine triphosphate (ATP) acts as a crucial energy currency in vivo , and it is a widely used energy and/or phosphate donor for enzyme-catalyzed reactions in vitro. In this study, we established an in vitro multi-enzyme cascade system for ATP production. Using adenosine and inorganic polyphosphate (polyP) as key substrates, we combined adenosine kinase and two functionally distinct polyphosphate kinases (PPKs) in a one-pot reaction to achieve chain-like ATP regeneration and production. Several sources of PPK were screened and characterized, and two suitable PPKs were selected to achieve high rates of ATP production. Among these, Sulfurovum lithotrophicum PPK (SlPPK) exhibited excellent activity over a wide pH range (pH 4.0–9.0) and synthesized ATP from ADP using short-chain polyP. Furthermore, it had a half-life > 155.6 h at 45 °C. After optimizing the reaction conditions, we finally carried out the coupling-catalyzed reaction with different initial adenosine concentrations of 10, 20, and 30 mM. The highest yields of ATP were 76.0, 70.5, and 61.3%, respectively. Graphical Abstract
研究领域:
生物化学
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生物技术
·
酶学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Kai-Yue Tan,Chun-Yan Li,Yong-Fei Li,Junjie Fei,Bin Yang,Ya-Jun Fu,Fang Li
Adenosine triphosphate (ATP) is mainly produced in the mitochondrion and used as a universal energy source for various cellular events. Various fluorescent probes for ATP have been established successfully, but most of them are not appropriate for monitoring the fluctuation of the mitochondrial ATP level. Herein, a fluorescent probe named Mito-Rh is first synthesized and used to recognize ATP in mitochondrion. In the probe, rhodamine, diethylenetriamine, and triphenylphosphonium are selected as fluorophore, reaction site, and mitochondrion-targeting group, respectively. Probe Mito-Rh shows high sensitivity to ATP with 81-fold fluorescence enhancement, and the detection range (0.1–10 mM) can match the concentration level of ATP in the mitochondrion. Moreover, Mito-Rh provides excellent selectivity toward ATP over other biological anions (ADP, AMP, GTP, CTP, UTP) owing to a concurrent effect of dual recognition sites (hydrogen bond and π–π stacking). In particular, the probe can localize in mitochondrion specifically and demonstrates utility in the real-time detection of mitochondrial ATP concentration changes.
研究领域:
细胞生物学
·
生物化学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Shun Hu,Xiaona Li,Kai Wang,Qiuhua Wu,Guolin Zhang,Xue Liu
A simple indicator displacement assay (IDA)-based array system was established by using Fe(III)-responsive fluorescent carbon dots (CDs) and Fe(III) to detect ATP-related physiological phosphates (PPs) and monitor ATP hydrolysis. The analysis was based on principal component analysis of differential CDs fluorescence regulated by Fe(III)/PPs complexes. By two modeling methods, two sensor arrays were established. That is, three CDs/one Fe(III) concentration (Array1) and one CD/four Fe(III) concentrations (Array2). Compared with Array1, Array2 saved a lot of trouble in preparing the CD sensors and the analytical performance it is hardly affected. The Array2 can distinguish PPs in serum environment and from interfering substances, such as amino acids and metal ions. Besides, quantitative analysis of ATP single-component system and qualitative analysis of ATP/ADP two-component system can be achieved. On this basis, ATP hydrolysis processes were monitored, and results suggested that ATP hydrolysis completed within 20 min under the catalysis of 24 U/mL apyrase. As ATP hydrolysis works for 30 min, its main products were mainly AMP and orthophosphate (Pi).
研究领域:
生物化学
·
分析化学
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产品关联
DOI
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2019
·Vol. 166
·期 15
·H777
·
IF 3.12
Wenwen Xu,Yonghong Sun,Cuimin Shao,Jianguo Wang
ATP, ADP, and cytidine triphosphate (CTP) are common sources of energy for various cellular activities. Therefore, a method that efficiently measures ATP, ADP, and CTP levels is urgently needed. In unbuffered solution, ATP, ADP, and CTP were observed to cause a new cathodic peak at a positive potential during benzoquinone (BQ) reduction. The hydrolysis of ATP, ADP, and CTP generated additional proton donors, which provided numerous protons for the reduction of BQ and formation of a large new cathodic peak. The peak current difference between the peaks caused by ATP, ADP, and CTP before and after hydrolysis was linear to the ATP, ADP, and CTP concentrations but independent of the BQ concentration. An electrochemical method for the detection of ATP, ADP, and CTP was developed using a bare glassy carbon electrode. The proposed method showed good selectivity for the detection of ATP samples.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Xiaomeng Zhou,Saijin Huang,Dan Zhang,Wenfeng Liu,Wenxing Gao,Yumeng Xue,Li Shang
Adenosine triphosphate (ATP) participates in the regulation of most biological processes, and the ATP level is closely associated with many diseases. However, it still remains challenging to achieve on-site monitoring of ATP in an equipment-free and efficient way. Microneedles, a minimally invasive technology that can extract biomarkers from liquid biopsies, have recently emerged as useful tools for early diagnosis of a broad range of diseases. In this work, we developed hydrogel microneedles that are loaded with ATP-specific dual-emitting gold nanoclusters (RhE-AuNCs) for fast sampling and on-needle detection of ATP. These RhE-AuNCs were photo-crosslinked to the hydrogel matrix to form a fluorescent microneedle patch. Based on the ATP-induced Förster resonance energy transfer in RhE-AuNCs, a highly selective, sensitive, and reliable ATP sensor was developed. Moreover, simultaneous capture and visual detection of ATP was achieved by the AuNC-loaded microneedle sensing platform, which exhibits promising sensing performance. This work provides a new approach to design a point-of-care ATP sensing platform, which also holds great potential for the further development of microneedle-based analytical devices.
研究领域:
生物医学工程
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分析化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 7
·1-11
·
IF 5.3
Liang Pengda,Lv Bei,Chen Ke,Li Dawei
The combination of CRISPR/Cas12a and functional DNA provides the possibility of constructing biosensors for detecting non-nucleic-acid targets. In the current study, the duplex protospacer adjacent motif (PAM) in the activator of CRISPR/Cas12a was used as a molecular switch, and a sensitive adenosine triphosphate (ATP) detection biosensor was constructed using an allosteric probe-conjugated PAM site formation in hybridization chain reaction (HCR) integrated with the CRISPR/Cas12a system (APF-CRISPR). In the absence of ATP, an aptamer-containing probe (AP) is in a stem-loop structure, which blocks the initiation of HCR. In the presence of ATP, the structure of AP is changed upon ATP binding, resulting in the release of the HCR trigger strand and the production of long duplex DNA with many PAM sites. Since the presence of a duplex PAM site is crucial for triggering the cleavage activity of CRISPR/Cas12a, the ATP-dependent formation of the PAM site in HCR products can initiate the FQ-reporter cleavage, allowing ATP quantification by measuring the fluorescent signals. By optimizing the sequence elements and detection conditions, the aptasensor demonstrated superior detection performance. The limit of detection (LOD) of the assay was estimated to be 1.16 nM, where the standard deviation of the blank was calculated based on six repeated measurements. The dynamic range of the detection was 25–750 nM, and the whole workflow of the assay was approximately 60 min. In addition, the reliability and practicability of the aptasensor were validated by comparing it with a commercially available chemiluminescence kit for ATP detection in serum. Due to its high sensitivity, specificity, and reliable performance, the APF-CRISPR holds great potential in bioanalytical studies for ATP detection. In addition, we have provided a proof-of-principle for constructing a CRISPR/Cas12a-based aptasensor, in which the PAM is utilized to regulate Cas12a cleavage activity. Graphical Abstract
研究领域:
生物化学
·
分子生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Qi Liu,Shushu Ding,Rui Gao,Guoyue Shi,Anwei Zhu
ATP-sensitive potassium (K ATP ) channels couple intracellular metabolism to the electrical activity by regulating K + flux across the plasma membrane, thus playing an important role in both normal and pathophysiology. To understand the mechanism of ATP regulating biological ion channels, developing an ATP-responsive artificial nanochannel is an appealing but challenging topic because K ATP channel is a heteromultimer of two subunits (potassium channel subunit (Kir6.x) and sulfonylurea receptor (SUR)) and exhibit dynamic functions with adjustability and reversibility. Inspired by the structure of K ATP channels, we designed a smart copolymer modified nanochannel that may address the challenge. In the tricomponent poly( N -isopropylacrylamide) (PNIPAAm, PNI)-based copolymer system, phenylthiourea was used to bind the phosphate units of nucleotides and phenylboronic acid was introduced to combine the pentose ring of the nucleoside unit. Besides, a –COOH group with electron-withdrawing property was added into the phenylthiourea units, which may promote the hydrogen-bond-donating ability of thiourea . Specially, the smart copolymer not only provided static binding sites for recognition but also translated the recognition of ATP into their dynamic conformational transitions by changing the hydrogen-bonding environments surrounding PNIPAAm chains, thus achieving the gating function of nanochannel, which resembled the integration and coordination of Kir6.x and SUR units in active K ATP . The ATP-regulated ion channel exhibited excellent stability and reversibility. This study is the first example showing how to learn from nature to assemble the ATP-responsive artificial nanochannel and demonstrate the possible mechanism of ATP gating.
研究领域:
生物化学
·
材料科学
·
分子生物学
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DOI
高影响因子
ACS Chemical Neuroscience
·
2019
·
IF 3.861
Lu Kou,Yuemei Duan,Pengcheng Wang,Yaru Fu,Nerek Darabedian,Yonghui He,Dianlu Jiang,Dinglong Chen,Juan Xiang,Guokun Liu,Feimeng Zhou
The aberrant autoxidation of norepinephrine (NE) in the presence of oxygen, which is accelerated by Fe(III), has been linked to the pathogenesis of the Parkinson’s disease (PD). Adenosine triphosphate (ATP), as a neurotransmitter whose release can be stimulated by tissue damage and oxidative stress, is co-stored and often co-released with NE in presynaptic terminals. We have shown previously that ATP inhibits the iron-catalyzed dopamine oxidation, thereby decreasing the production of certain neurotoxins such as 6-hydroxydopamine. Whether ATP plays a similar role in Fe(III)-catalyzed NE oxidation and how it maintains the NE stability have not been investigated. Here, we studied the coordination in a ternary complex among NE, Fe(III), and ATP, and found that Fe(III) is coordinated as a octahedral center by NE and ATP. Voltammetry and mass spectrometry were employed to examine this ternary complex’s modulation of the NE autoxidation. NE–Fe(III)–ATP plays a protective role to modulate the autoxidation and Fe(III)-catalyzed oxidation of NE. The ternary complex can be detected in the substantia nigra (SN), locus coeruleus (LC), and striatum regions of C57BL/6 wild-type mice. In contrast, the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse brains displayed a significant decrease of the ternary complex in the SN region and an increase in the LC and striatum areas. We posit that the ternary complex is produced by noradrenergic neurons as a protective regulator against neuronal damage and oxidative stress, contributing to the lower vulnerability of LC neurons with respect to that of SN neurons.
研究领域:
神经科学
·
医学
·
生物化学
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DOI
PubMed
高影响因子
TALANTA
·
2015
·
IF 3.545
Yakun Lian,Hua Jiang,Jinzhou Feng,Xiaoyan Wang,Xiandeng Hou,Pengchi Deng
ATP , ADP and AMP are energy substances with vital biological significance. Based on the structural differences, a simple, rapid and comprehensive method has been established by 1 H and 31 P Nuclear Magnetic Resonance ( 1 H-NMR and 31 P-NMR) spectroscopies. Sodium 3-(trimethylsilyl) propionate-2,2,3,3-d 4 (TMSP) and anhydrous disodium hydrogen phosphate (Na 2 HPO 4 ) were selected as internal standards for 1 H-NMR and 31 P-NMR, respectively. Those three compounds and corresponding internal standards can be easily distinguished both by 1 H-NMR and 31 P-NMR. In addition, they all have perfect linearity in a certain range: 0.1–100 mM for 1 H-NMR and 1–75 mM for 31 P-NMR. To validate the precision of this method, mixed samples of different concentrations were measured. Recovery experiments were conducted in serum (91–113% by 1 H-NMR and 89–113% by 31 P-NMR).
研究领域:
生物化学
·
医学检测
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DOI
PubMed
高影响因子
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