AMINO ACIDS
·
2023
·Vol. 55
·期 4
·499-507
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IF 3.5
Qin Tiansheng,Huang Xiande,Zhang Qianqian,Chen Fan,Zhu Jiaojiao,Ding Yaoyao
During trauma and surgery, bleeding is a major concern. One of the crucial strategies for hemostasis is the use of biological hemostatic material. Herein, we reported an amino acid-based hydrogel FmocF-ADP hydrogel, which consisted of N-[(9H-fluoren-9-ylmethoxy) carbonyl]-3-phenyl-L-alanine (FmocF) and adenosine diphosphate (ADP) sodium solution. The hydrogel was created by FmocF self-assembling to nanofiber in ADP sodium solution and then cross-linking to hydrogel. FmocF-ADP hydrogel showed good in vitro coagulation activity as measured by whole blood clotting assays, platelet clotting assays, platelet activation assays, and platelet adhesion assays. Further, it was noted to reveal an exceptional in vivo hemostatic effect in a mouse liver bleeding model. Together with the previous report of the good biocompatibility and antimicrobial activity of FmocF hydrogel, our study would extend the biomedical application of FmocF hydrogel. In conclusion, the present study would provide a constructive strategy for the development of new antimicrobial and hemostatic materials or develop a potential hemostatic material.
研究领域:
生物医学工程
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材料科学
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止血
·
药剂学
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DOI
PubMed
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
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
ANALYST
·
2018
·Vol. 143
·期 11
·2501-2507
·
IF 3.864
Yong Liu,Xiaolin Xu,Haitang Yang,Ensheng Xu,Shuangshuang Wu,Wei Wei,Jin Chen
Poly(ADP-ribose) polymerase-1 (PARP-1) is a highly conserved nuclear enzyme, which binds tightly to damaged DNA and plays a key role in DNA repair, recombination, proliferation, and genomic stability. However, due to the poor electrochemical and optical activity of PARP-1 and its product PAR, only a few studies on its activity detection method have been reported. Herein, we report a simple and sensitive colorimetric strategy to monitor PARP-1 activity based on enzyme-initiated auto-PARylation-controlled aggregation of hemin-graphene nanocomposites (H-GNs). PARP, activated by dsDNA, catalyzed its substrate nicotinamide adenine dinucleotide (NAD+) to polymerize as a poly(ADP-ribose) polymer (PAR). PAR possesses several negative charges, and its charge density is twice that of a single-stranded DNA, which greatly impacts the dispersibility of H-GNs; due to their peroxidase-like catalytic activities, H-GNs can catalyze the chromogenic reaction of TMB and H2O2. As a result, in the presence of different PARP-1 activities, the supernatant of the corresponding solution contained different amounts of dispersed H-GNs and showed different colors after the chromogenic reaction that could be discerned easily by the absorbance or the color changes of the solution. The method was simple, sensitive, and reliable. The proposed method displays a linear range from 0.05 to 1 U with a detection limit of 0.03 U. In addition, this new method has been successfully applied to detect PARP-1 activity in human serum and different cancer cells and evaluate PARP-1 inhibitors.
研究领域:
生物化学
·
分子生物学
·
癌症研究
·
纳米技术
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DOI
PubMed
高影响因子
Biosensors-Basel
·
2023
·Vol. 13
·期 7
·723
·
IF 5.4
Fengli Gao,Gang Liu,Yishu Qiao,Xiuwen Dong,Lin Liu
This work reports the development of a fluorescence method for the detection of poly(ADP-ribose) polymerase-1 (PARP1), in which a phenylboronic acid-modified fluorescein isothiocyanate dye (FITC-PBA) was used to recognize the formed poly(ADP-ribose) (PAR) polymer. The detection system was designed by conjugating recombinant streptavidin (rSA) with PARP1-specific double-stranded DNA (dsDNA) through streptavidin–biotin interaction. Capture of PARP1 via rSA–biotin–dsDNA allowed for the poly-ADP-ribosylation (PARylation) of both rSA and PARP1 in a homogeneous solution. The resulting rSA–biotin–dsDNA/PAR conjugates were then captured and separated via the commercialized nitrilotriacetic acid–nickel ion-modified magnetic bead (MB-NTA-Ni) through the interaction between NTA–Ni on MB surface and oligohistidine (His6) tag in rSA. The PAR polymer could capture the dye of FITC-PBA through the borate ester interaction between the boronic acid moiety in PBA and thecis-diol group in ribose, thus causing a decrease in fluorescence signal. The PARylation of streptavidin and the influence of steric hindrance on PARylation efficiency were confirmed using reasonable detection strategies. The method showed a wide linear range (0.01~20 U) and a low detection limit (0.01 U). This work should be valuable for the development of novel biosensors for the detection of poly(ADP-ribose) polymerases and diol-containing species.Keywords:streptavidin;nitrilotriacetic acid;magnetic bead;poly(ADP-ribose) polymerase;boronic acid;immobilization-free
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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生物技术
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DOI
PubMed
高影响因子
JOURNAL OF BIOLOGICAL CHEMISTRY
·
2025
·
IF 3.9
Bao-Di Ma,Jia-Yi Li,Jian-He Xu,Tao Yu,Xu-Dong Kong
Methanol dehydrogenase (MDH), a representative of Type III alcohol dehydrogenases (ADHs), plays a pivotal role in methanol assimilation pathways, making it a key enzyme for the biosynthesis of chemicals and fuels from one-carbon feedstocks. An activator protein belonging to the Nudix hydrolase family, ACT, was found to increase the activity of MDH by 40-fold. Despite the widespread observation of this in vitro activation phenomenon in pairs of type III alcohol dehydrogenases and Nudix hydrolases, the mechanistic details have remained unresolved for decades. Here, we uncover a regulation mechanism in which MDH activation arises from the hydrolytic removal of ADP-ribose (ADPR), a potent inhibitor derived from NAD + degradation, by the ADPRase activity of ACT. This discovery challenges the previously proposed 'activation' models, revealing that ACT-mediated ADPR clearance disinhibits MDH rather than directly enhancing catalysis. By combining crystallographic analysis, kinetics, and inhibition assays, we demonstrate that ADPR inhibits MDHs with submicromolar K i values, highlighting its potential regulatory role in metabolic networks. Our findings redefine the widespread ‘activation’ of type III ADHs, providing valuable insights into alcohol metabolism and new directions for engineering synthetic methanol utilization pathways.
研究领域:
生物化学
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结构生物学
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酶学
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代谢工程
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微生物学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Ensheng Xu,Haitang Yang,Lina Wu,Jin Chen,Wei Wei,Yong Liu,Songqin Liu
Since the expression of poly(ADP-ribose) polymerase-1 (PARP-1) was significantly up-regulated in several primary malignancies, it was important to construct a sensitive and rapid PARP-1 assay for early diagnosis of cancer. When PARP-1 was activated by active dsDNA, it generates a number of negatively charged poly(ADP-ribose) (PAR) polymers, which can be readily detected. Here, we developed a label-free electrochemical (EC) biosensor for PARP-1 activity based on the electrostatic attraction between negatively charged PAR and positively charged amino-functioned mesoporous silica films (NH 2 -MSFs). Negatively charged PAR was absorbed on the NH 2 -MSFs surface and suppressed the electrochemical signal as the PAR prevented the electroactive probe reaching to the ITO electrode. The electrochemical signal for PAR linearly decreased from 0.01 U to 1.2 U with a detection limit of 0.005 U, which is comparable to the PARP-1 biosensors reported before. The strategy has been used to evaluate PARP-1 inhibitors and to detect PARP-1 activity in human serum and real cancer cells lysate with satisfactory results, indicating that it was a promising candidate for point-of-care diagnostics.
研究领域:
生物化学
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医学诊断
·
材料科学
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2016
·
IF 7.145
Yuanyuan Xu,Li Liu,Zhaoyin Wang,Zhihui Dai
A stable and reusable electrochemical biosensor for the label-free detection of poly(ADP-ribose) polymerase (PARP) is designed in this work. C-kit-1, a thiol-modified G-quadruplex oligonucleotide, is first self-assembled on a gold electrode surface. The G-quadruplex structure of c-kit-1 can specifically tether and activate PARP, resulting in the generation of negatively charged poly(ADP-ribose) polymer (PAR). On the basis of electrostatic attraction, PAR facilitates the surface accumulation of positively charged electrochemical signal molecules. Through the characterization of electrochemical signal molecules, the label-free quantification of PARP is simply implemented. On the basis of the proposed method, selective quantification of PARP can be achieved over the linear range from 0.01 to 1 U with a calculated detection limit of 0.003U. Further studies also demonstrate the applicability of the proposed method to biosamples revealing the broad potential in practical applications. Furthermore, inhibitor of PARP has also been detected with this biosensor. Meanwhile, benefited from self-assembly on solid surface, this biosensor possesses two important features, i.e., reusability and stability, which are desirable in related biosensors.
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
TALANTA
·
2024
·
IF 5.6
Suling Yang,Yaliang Huang,Tingting Yang,Jingze Li,Jiaxin Tian,Lin Liu
Poly(ADP-ribose)polymerase-1 (PARP1) could be activated by binding to nucleic acids with specific sequences, thus catalyzing the poly-ADP-ribosylation (PARylation) of target proteins including PARP1 itself. Most of the previously reported electrochemical methods for the determination of PARP1 were relied on the electrostatic interactions, which required the pre-immobilization of DNA on an electrode for the capture of PARP1. Herein, we reported an “immobilization−free” electrochemical strategy for the assays of PARP1 on the basic of avidin-biotin interaction. Once PARP1 was activated by binding with the specific double-stranded DNA (dsDNA) in a homogeneous solution, the biotinylated nicotinamide adenine dinucleotide (biotin-NAD + ) was transferred onto PARP1, resulting in the formation of biotinylated PAR polymers. The resulting biotinylated PAR polymers were then captured by a neutravidin (NA)-modified electrode through avidin-biotin interactions. The rich biotin moieties in the PAR polymers allowed for the capture of NA-modified silver nanoparticles (NA-AgNPs) through the avidin-biotin interactions. The surface-tethered AgNPs produced a well-defined electrochemical signal due to the characteristic solid-state Ag/AgCl process. The “immobilization−free”, electrostatic interaction-independent electrochemical biosensor exhibited low background current, high sensitivity, and good stability. It has achieved the determination of PARP1 with a detection limit down to 0.7 mU. The biosensor was further applied to determine the inhibition efficiency of potential inhibitors with a satisfactory result. This method shows promising potential applications in PARP1-related clinical diagnosis and drug discovery.
研究领域:
生物化学
·
医学诊断
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Ning Xia,Daohong Wu,Ting Sun,Yilin Wang,Xiaoyu Ren,Feng Zhao,Lin Liu,Xinyao Yi
Poly(ADP-ribose) polymerase-1 (PARP-1), as a highly conserved nuclear enzyme, can catalyze the transfer of multiple ADP-ribose units from β-nicotinamide adenine dinucleotide (NAD + ) to substrate proteins, including PARP-1 itself. In this work, two boronic acid derivatives, ferroceneboronic acid (FcBA) and 4-mercaptophenylboronic acid (MPBA) were used as the signal probes for electrochemical and colorimetric detection of PARP-1, respectively. The detection platform was constructed by modification of double-stranded DNA (dsDNA) on the surface of magnetic beads (MBs). Once PARP-1 was captured by the MB-dsDNA, auto-PARylation was initiated and the PAR products consisting of abundant ribose units were covalently bound to FcBA or MPBA through the formation of boronate ester covalent bonds. Sequestration of FcBA or MPBA by the PAR-covered MBs led to the decrease in the electrochemical signal of FcBA or preventing the MPBA-triggered aggregation of gold nanoparticle indicators. The change of oxidation current or adsorption intensity (color) was dependent upon the concentration and activity of PARP-1. A wide linear range of 0.1 ∼ 50 U was obtained for the electrochemical or colorimetric assays. The methods have been used to evaluate the inhibition efficiency and monitor the PARP-1 activity in living and apoptotic cancer cells.
研究领域:
生物化学
·
分析化学
·
分子生物学
·
癌症研究
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DOI
高影响因子
IMMUNOLOGICAL INVESTIGATIONS
·
2023
·
IF 2.8
Dongdong Dou,Meirong Bi,Xiuyun Li,Nan Zhang,Mi Xu,Aili Guo,Feng Li,Weiwei Zhu
Background Childhood asthma is a major global health concern. ADP-ribosylation factor 6 (ARF6) is a low-molecular-weight GTPase; however, its role in childhood asthma remains unclear.Methods Ovalbumin (OVA)-challenged neonatal mice and transforming growth factor-β1 (TGF-β1)-induced BEAS-2B cells were used as in vivo and in vitro models of childhood asthma, respectively.Results Upon OVA stimulation, ARF6 expression was upregulated in the lung tissue. Neonatal mice administered SehinH3 (an ARF6 inhibitor) exhibited improved pulmonary pathological injury, along with reduced inflammatory cell infiltration in the lungs and cytokine release in bronchial alveolar lavage fluid and serum (interleukin [IL]−3, IL−5, IL−13, IgE, and OVA-specific IgE). SehinH3 treatment restrained epithelial – mesenchymal transition (EMT) in the lungs of asthmatic mice, as evidenced by increased E-cadherin and decreased N-cadherin and α-smooth muscle actin expression. Different TGF-β1 exposures to BEAS-2B cells induced a time- and dose-dependent increase in ARF6 expression in vitro. Upon TGF-β1 stimulation, ARF6 knockdown repressed EMT and SehinH3 treatment caused similar results in BEAS-2B cells. The transcription factor E2F8 is involved in diverse biological functions and its increased expression was confirmed in vivo and in vitro. Dual-luciferase assays confirmed that E2F8 binds to the ARF6 promoter and promotes its transcriptional activity. In vitro results revealed that E2F8 silencing suppressed EMT, whereas rescue experiments showed that ARF6 overexpression partly reversed these phenomena.Conclusion Our study showed that ARF6 is associated with childhood asthma progression and may be positively regulated by E2F8. These results provide insight into the pathogenesis and treatment of childhood asthma.
研究领域:
免疫学
·
呼吸病学
·
分子生物学
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL RESEARCH IN TOXICOLOGY
·
2018
·
IF 3.432
Wenjing Dong,Bingwei Yang,Yawen Wang,Jia Yuan,Yunqi Fan,Erqun Song,Yang Song
Polybrominated diphenyl ethers (PBDEs) are emerging organic environmental pollutants, which were accused of various toxic effects. Here, we studied the role of a potential PBDEs quinone metabolite, PBDEQ, on cytotoxicity, oxidative DNA damage, and the alterations of signal cascade in HeLa cells. PBDEQ exposure leads to reactive oxygen species (ROS) accumulation, mitochondrial membrane potential (MMP) loss, lactate dehydrogenase (LDH) release, increasing terminal transferase-mediated dUTP-biotin nick end labeling (TUNEL) positive foci, and the elevation of apoptosis rate. Furthermore, we showed PBDEQ exposure result in increased DNA migration, micronucleus frequency, and the promotion of 8-OHdG and phosphorylation of histone H2AX (γ-H2AX) levels. Mechanism study indicated that PBDEQ caused poly(ADP-ribose) polymerase 1 (PARP-1) activation and apoptosis-inducing factor (AIF) nuclear translocation. All together, these results confirmed the occurrence of parthanatos-like cell death upon PBDEQ exposure.
研究领域:
毒理学
·
细胞生物学
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环境科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）