MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 3
·1-8
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IF 6.232
Chen Yan,Yin Huanshun,Sui Chengji,Fu Weizhang,Yang Yue,Ai Shiyun
A photoelectrochemical method was proposed for DNA hydroxymethylation determination using black TiO 2 (B-TiO 2 ) nanosphere as photoactive material and ZnO as photoactivity inhibitor. After hydroxymethylated DNA (5hmC-DNA) was captured on the probe modified B-TiO 2 /ITO electrode surface through hybridization, a glycosyl can be then transferred from uridine diphosphoglucose to 5hmC-DNA and formed a covalent structure with –CH 2 OH in the presence of T4 β-glucosyltransferase (β-GT). Afterwards, based on a series of covalent reaction, amino functionalized ZnO nanoparticles are further immobilized to the surface of the electrode. Due to the capacity to expend the irradiation light and the photogenerated electron of electron donor, the modified ZnO nanoparticles can result in a decreased photocurrent. The developed method shows wide linear ranges from 0.05-200 nM for hydroxymethylated DNA and 1-220 unit·mL −1 for T4-β-glucosyltransferase. The corresponding determination limits were 0.013 nM and 0.24 unit·mL −1 , respectively. The enzyme activity inhibited by 4-phenylimidazole was evaluated. This photoelectrochemical method shows high specificity for 5hmC-DNA (compared to 5fC, 5mC, m6A, control) and β-GT (compared to β-AGT, UGT2B7), and shows excellent stability for testing 5hmC (RSD = 2.75%).
Schematic representation of photoelectrochemical method for DNA hydroxymethylation and β-glucosyltransferase detection based on the glycosylation reaction of –CH 2 OH in 5-hydroxymethylcytosine and the inhibition activity of ZnO to the photoactivity of black TiO 2 nanospheres.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Materials Today Chemistry
·
2022
·
IF 7.613
Yulin Zheng,Yunlei Zhou,Xiaoting Cui,Huanshun Yin,Shiyun Ai
The photoactivity of CdS nanorods was greatly improved by amino functionalized accordion-like MXene and spherical ZnSnO 3 . MXene possesses good electron transfer capability and ZnSnO 3 presents matched energy band with CdS, which deeply accelerate the electron transfer and prevent the recombination of photogenerated electron-hole pair, leading to a strong photoelectrochemical (PEC) response. Taking the merit of the improved photoactivity of CdS nanorods, a novel PEC biosensor was constructed for DNA hydromethylation detection based on immune recognition of target molecule, where 5-hydroxymethyl-2′-deoxycytidine triphosphate (5hmdCTP) was employed as detect target, CdS/MXene was used as photoactive material, and ZnSnO 3 was adopted as signal amplification unit. Under enzymatic covalent reaction of –CH 2 OH of 5hmdCTP with –NH 2 of MXene, 5hmdCTP was specifically recognized and captured. Then, taking advantages of the covalent reaction between phosphate group of 5hmdCTP and ZnSnO 3 , the signal amplification unit was captured. Under the optimum conditions, this PEC biosensor presents wide linear range of 0.008–100 nM and low detection limit of 4.21 pM (3σ). The applicability of the developed method was evaluated by investigating the effect of Cd 2+ and perfluorohexane compound pollutant on 5-hydroxymethylcytosine content in the genomic DNA of the roots and leaves of wheat seedlings.
研究领域:
生物传感器
·
环境科学
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
ELECTROCHIMICA ACTA
·
2015
·
IF 4.504
Yunlei Zhou,Zhiqing Yang,Xue Li,Yue Wang,Huanshun Yin,Shiyun Ai
DNA hydroxymethylation (5-hydroxymethylcytosine, 5hmC) is a kind of new epigenetic modification, which plays key roles in nuclear reprogramming, regulates the gene activity, and initiates the DNA demethylation in mammals. For further understanding the functions of 5hmC and the correlation with tumour, it is essential to develop sensitive and selective methods for detecting and sequencing 5hmC. Herein, a kind of electrochemical biosensor was fabricated for 5hmC detection based on the glycosylation modification of 5hmC and enzymatic signal amplification. Under the catalytic effect of T4 β-glucosyltransferase, the 5hmC in target DNA was chemically modified with glucose. Then with the bridge connection of 1,4-phenyldiboronic acid, alkaline phosphatase was further captured on the electrode surface to catalyze the hydrolysis of p -nitrophenyl phosphate disodium salt to produce p -nitrophenol. Based on the relationship between the electrochemical oxidation signal of p -nitrophenol and the concentration of target DNA, the 5hmC level can be detected with high sensitivity and selectivity. The developed method also showed excellent reproducibility and stability.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
表观遗传学
✓ 已验证
产品关联
DOI
高影响因子
Redox Biology
·
2022
·
IF 10.787
Qian-Qian Han,Peng-Fei Wu,Yi-Heng Li,Yu Cao,Jian-Guo Chen,Fang Wang
Vitamin C, a key antioxidant in the central nervous system, cycles between ascorbic acid and dehydroascorbic acid under pathophysiological conditions. Clinical evidence supports that the absence of vitamin C may be linked to depressive symptoms, but much less is known about the mechanism. Herein, we show that chronic stress disrupts the expression of ascorbic acid transporter, sodium-dependent vitamin C transport 2, and induces a deficiency in endogenous ascorbic acid in the medial prefrontal cortex, leading to depressive-like behaviors by disturbing redox-dependent DNA methylation reprogramming. Attractively, ascorbic acid (100 mg/kg-1000 mg/kg, intraperitoneal injection, as bioequivalent of an intravenous drip dose of 0.48 g–4.8 g ascorbic acid per day in humans) produces rapid-acting antidepressant effects via triggering DNA demethylation catalyzed by ten-eleven translocation dioxygenases. In particular, the mechanistic studies by both transcriptome sequencing and methylation sequencing have shown that S100 calcium binding protein A4, a potentially protective factor against oxidative stress and brain injury, mediates the antidepressant activity of ascorbic acid via activating erb-b2 receptor tyrosine kinase 4 (ErbB4)-brain derived neurotrophic factor (BDNF) signaling pathway. Overall, our findings reveal a novel nutritional mechanism that couples stress to aberrant DNA methylation underlying depressive-like behaviors. Therefore, application of vitamin C may be a potential strategy for the treatment of depression.
研究领域:
神经科学
·
药理学
·
分子生物学
·
精神病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Jia Ding,Fei Liu,Chunling Qi,Yunlei Zhou,Huanshun Yin,Shiyun Ai
A novel photoelectrochemical (PEC) biosensor was constructed for DNA hydroxymethylation detection based on Bi 4 NbO 8 Cl/PTC-NH 2 heterojunction and Fe-MIL-88NH 2 , where Bi 4 NbO 8 Cl/PTC-NH 2 was employed as photoactive material and Fe-MIL-88NH 2 was adopted as artificial hydrogen peroxide mimic enzyme. Based on the structure of hydroxymethylated cytosine in DNA sequence, 5-hydroxymethyl-2’-deoxycytidine (5hmdC) was employed as detected target molecule for the first time using PEC technique. After Bi 4 NbO 8 Cl/PTC-NH 2 heterojunction was modified on the substrate electrode, thioglycolic acid (TGA) was captured by forming amide bond, where the sulfhydryl group (-SH) of TGA was employed as the specific recognition of 5hmdC. Under M. HhaI catalysis, the −CH 2 OH of 5hmdC reacted selectively with the –SH. Finally, Fe-MIL-88NH 2 was modified on the electrode based on the crosslinker of 4-formylphenylboronic acid (FPBA), where FPBA possessed two active groups of formyl and boronic acid group, reacting with glycosyl of 5hmdC and amino of Fe-MIL-88NH 2 . Under the catalysis effect of Fe-MIL-88NH 2 , the insoluble substance was produced on the surface of electrodes, resulting in distinct lessening of PEC response. Under optimal conditions, the photocurrent shows a linear relationship with the logarithm value of 5hmdC concentration from 0.01-100 nM with a low detection limit of 6.48 pM. More importantly, this method can be applied to investigate the influence of exogenous phytohormone on 5hmdC content in the genomic DNA of rice seedling roots and leaves.
研究领域:
材料科学
·
分析化学
·
生物传感器
·
生物化学
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纳米技术
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电化学
·
基因组学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Jia Ding,Lanlan Gao,Lulu Cao,Yunlei Zhou,Hao Wang,Jingrui Duan,Huanshun Yin,Jianchao Sun,Shiyun Ai
DNA hydroxymethylation is an important epigenetic modification, which plays crucial roles in many biological processes, such as gene expression and cell differentiation. Herein, a novel photoelectrochemical immunosensor was built for detecting 5-hydroxymethylcytosine (5hmC), where the perovskite Bi 4 TaO 8 Cl sensitized by 3,4,9,10-perylenetetracarboxylicacid (PTCA) was implemented as photoactive material, anti-5hmC antibody was served as target recognition reagent, ZrO 2 was used for connecting more modifications, and ferrocene was adopted as signal amplification unit. The prepared nanomaterials of Bi 4 TaO 8 Cl, PTCA and ZrO 2 were well characterized. The possible mechanism for the improved PEC response of Bi 4 TaO 8 Cl sensitized by PTCA was investigated carefully, indicated that the matched energy band facilitated the migration of photogenerated electron. In addition, the possible mechanism for signal amplification caused by ferrocene was investigated, showing that ferrocene acted as electron donor to accelerate electron transfer. Under the optimal condition, the biosensor manifested wide detection range from 0.02 to 100 nM with the low detection limit of 5.2 pM (S/N = 3). Moreover, the applicability of the biosensor was demonstrated by investigating the effect of heavy metal Cd 2+ and perfluorooctanoic acid on 5hmC content in the leaves and roots of wheat seedlings, as well as the effect of ALV-J on 5hmC content in chicken serum.
研究领域:
生物化学
·
分析化学
·
材料科学
·
环境科学
·
分子生物学
✓ 已验证
DOI
高影响因子
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