Co-reporter:Kunli Zhao;Yu Tang;Zhen Wang;Juan Zhang;Chunyang Lei;Honghui Wang;Hao Li;Yan Huang;Shouzhuo Yao
Chemical Communications 2017 vol. 53(Issue 82) pp:11326-11329
Publication Date(Web):2017/10/12
DOI:10.1039/C7CC06833F
A versatile bio-logic system is proposed using a surface charge tunable fluorescent protein (H39GFP) as a generic processor. On the basis of the cell penetration of H39GFP triggered by H+ and metal ions, OR logic gates are successfully operated in living cells with transfection of functional nucleic acids (Cy5 labelled DNA and siRNA) as the readout, as well as RNA interference.
Co-reporter:Yong Li;Sujuan Sun;Lin Fan;Shanfang Hu; Yan Huang; Ke Zhang; Zhou Nie; Shouzhou Yao
Angewandte Chemie International Edition 2017 Volume 56(Issue 47) pp:14888-14892
Publication Date(Web):2017/11/20
DOI:10.1002/anie.201708327
AbstractA novel and versatile peptide-based bio-logic system capable of regulating cell function is developed using sortase A (SrtA), a peptide ligation enzyme, as a generic processor. By modular peptide design, we demonstrate that mammalian cells apoptosis can be programmed by peptide-based logic operations, including binary and combination gates (AND, INHIBIT, OR, and AND-INHIBIT), and a complex sequential logic circuit (multi-input keypad lock). Moreover, a proof-of-concept peptide regulatory circuit was developed to analyze the expression profile of cell-secreted protein biomarkers and trigger cancer-cell-specific apoptosis.
Co-reporter:Yong Li;Sujuan Sun;Lin Fan;Shanfang Hu; Yan Huang; Ke Zhang; Zhou Nie; Shouzhou Yao
Angewandte Chemie 2017 Volume 129(Issue 47) pp:15084-15088
Publication Date(Web):2017/11/20
DOI:10.1002/ange.201708327
AbstractA novel and versatile peptide-based bio-logic system capable of regulating cell function is developed using sortase A (SrtA), a peptide ligation enzyme, as a generic processor. By modular peptide design, we demonstrate that mammalian cells apoptosis can be programmed by peptide-based logic operations, including binary and combination gates (AND, INHIBIT, OR, and AND-INHIBIT), and a complex sequential logic circuit (multi-input keypad lock). Moreover, a proof-of-concept peptide regulatory circuit was developed to analyze the expression profile of cell-secreted protein biomarkers and trigger cancer-cell-specific apoptosis.
Co-reporter:Zhuoliang Liu, Xingyu Luo, Zhu Li, Yan Huang, Zhou NieHong-Hui Wang, Shouzhuo Yao
Analytical Chemistry 2017 Volume 89(Issue 3) pp:
Publication Date(Web):January 11, 2017
DOI:10.1021/acs.analchem.6b04360
Fluorogenic probes targeting G-quadruplex structures have emerged as the promising toolkit for functional research of G-quadruplex and biosensor development. However, their biosensing applications are still largely limited in in-tube detection. Herein, we proposed a fluorescent bioimaging method based on enzyme-generated G-quadruplexes for detecting apoptotic cells at the cell and tissue level, namely, terminal deoxynucleotidyl transferase (TdT)-activated de novo G-quadruplex synthesis (TAGS) assay. The detection target is genomic DNA fragmentation, a biochemical hallmark of apoptosis. The TAGS assay can efficiently “tag” DNA fragments via using their DNA double-strand breaks (DSBs) to initiate the de novo synthesis of G-quadruplexes by TdT with an unmodified G-rich dNTP pool, followed by a rapid fluorescent readout upon the binding of thioflavin T (ThT), a fluorogenic dye highly specific for G-quadruplex. The feasibility of the TAGS assay was proved by in situ sensitive detection of individual apoptotic cells in both cultured cells and tissue sections. The TAGS assay has notable advantages, including being label-free and having quick detection, high sensitivity and contrast, mix-and-read operation without tedious washing, and low cost. This method not only shows the feasibility of G-quadruplex in tissue bioanalysis but also provides a promising tool for basic research of apoptosis and drug evaluation for antitumor therapy.
Co-reporter:Xiaohua Zhu, Tingbi Zhao, Zhou Nie, Zhuang Miao, Yang Liu and Shouzhuo Yao
Nanoscale 2016 vol. 8(Issue 4) pp:2205-2211
Publication Date(Web):03 Dec 2015
DOI:10.1039/C5NR07826A
In this work, nitrogen-doped carbon nanoparticle (N-CNP) modulated turn-on fluorescent probes were developed for rapid and selective detection of histidine. The as synthesized N-CNPs exhibited high fluorescence quantum yield and excellent biocompatibility. The fluorescence of N-CNPs can be quenched selectively by Cu(II) ions with high efficiency, and restored by the addition of histidine owing to the competitive binding of Cu(II) ions and histidine that removes Cu(II) ions from the surface of the N-CNPs. Under the optimal conditions, a linear relationship between the increased fluorescence intensity of N-CNP/Cu(II) ion conjugates and the concentration of histidine was established in the range from 0.5 to 60 μM. The detection limit was as low as 150 nM (signal-to-noise ratio of 3). In addition, the as-prepared N-CNP/Cu(II) ion nanoprobes showed excellent biocompatibility and were applied for a histidine imaging assay in living cells, which presented great potential in the bio-labeling assay and clinical diagnostic applications.
Co-reporter:Xiaohua Zhu, Mengjia Liu, Yang Liu, Ruwen Chen, Zhou Nie, Jinghong Li and Shouzhuo Yao
Journal of Materials Chemistry A 2016 vol. 4(Issue 23) pp:8974-8977
Publication Date(Web):11 Apr 2016
DOI:10.1039/C6TA01923D
Carbon coated hollow mesoporous FeP microcubes derived from Prussian blue were superior in catalytic activity and durability toward electrochemical hydrogen evolution with an overpotential of 115 mV to drive 10 mA cm−2.
Co-reporter:Yong Li, Wang Li, Kai-Yu He, Pei Li, Yan Huang, Zhou Nie and Shou-Zhuo Yao
Nanoscale 2016 vol. 8(Issue 16) pp:8591-8599
Publication Date(Web):23 Mar 2016
DOI:10.1039/C6NR01072E
In natural biological systems, proteins exploit various functional peptide motifs to exert target response and activity switch, providing a functional and logic basis for complex cellular activities. Building biomimetic peptide-based bio-logic systems is highly intriguing but remains relatively unexplored due to limited logic recognition elements and complex signal outputs. In this proof-of-principle work, we attempted to address these problems by utilizing multi-functional peptide probes and the peptide-mediated nanoparticle assembly system. Here, the rationally designed peptide probes function as the dual-target responsive element specifically responsive to metal ions and enzymes as well as the mediator regulating the assembly of gold nanoparticles (AuNPs). Taking advantage of Zn2+ ions and chymotrypsin as the model inputs of metal ions and enzymes, respectively, we constructed the peptide logic system computed by the multi-functional peptide probes and outputted by the readable colour change of AuNPs. In this way, the representative binary basic logic gates (AND, OR, INHIBIT, NAND, IMPLICATION) have been achieved by delicately coding the peptide sequence, demonstrating the versatility of our logic system. Additionally, we demonstrated that the three-input combinational logic gate (INHIBIT-OR) could also be successfully integrated and applied as a multi-tasking biosensor for colorimetric detection of dual targets. This nanoparticle-based peptide logic system presents a valid strategy to illustrate peptide information processing and provides a practical platform for executing peptide computing or peptide-related multiplexing sensing, implying that the controllable nanomaterial assembly is a promising and potent methodology for the advancement of biomimetic bio-logic computation.
Co-reporter:Binbin Xiang, Kaiyu He, Rong Zhu, Zhuoliang Liu, Shu Zeng, Yan Huang, Zhou Nie, and Shouzhuo Yao
ACS Applied Materials & Interfaces 2016 Volume 8(Issue 35) pp:22801
Publication Date(Web):August 16, 2016
DOI:10.1021/acsami.6b03572
DNA hydrogel is a promising biomaterial for biological and medical applications due to its native biocompatibility and biodegradability. Herein, we provide a novel, versatile, and cost-effective approach for self-assembly of DNA hydrogel using the enzymatically polymerized DNA building blocks. The X-shaped DNA motif was elongated by terminal deoxynucleotidyl transferase (TdT) to form the building blocks, and hybridization between dual building blocks via their complementary TdT-polymerized DNA tails led to gel formation. TdT polymerization dramatically reduced the required amount of original DNA motifs, and the hybridization-mediated cross-linking of building blocks endows the gel with high mechanical strength. The DNA hydrogel can be applied for encapsulation and controllable release of protein cargos (for instance, green fluorescent protein) due to its enzymatic responsive properties. Moreover, this versatile strategy was extended to construct a functional DNAzyme hydrogel by integrating the peroxidase-mimicking DNAzyme into DNA motifs. Furthermore, a hybrid cascade enzymatic reaction system was constructed by coencapsulating glucose oxidase and β-galactosidase into DNAzyme hydrogel. This efficient cascade reaction provides not only a potential method for glucose/lactose detection by naked eye but also a promising modular platform for constructing a multiple enzyme or enzyme/DNAzyme hybrid system.Keywords: cascade reaction; DNA hydrogel; multiple-enzyme system; protein encapsulation; self-assembly
Co-reporter:Yong Li, Pei Li, Rong Zhu, Chao Luo, Hao Li, Shanfang Hu, Zhou Nie, Yan Huang, and Shouzhuo Yao
Analytical Chemistry 2016 Volume 88(Issue 22) pp:11184
Publication Date(Web):October 25, 2016
DOI:10.1021/acs.analchem.6b03389
Bioimaging probes for accurately monitoring apoptosis process have extensive significance for cell biological studies and clinical investigations. Herein, novel multifunctional peptide-tailored gold nanoclusters (AuNCs) have been developed for real-time imaging of caspase-indicated cell apoptosis. The AuNCs nanoprobe was facilely prepared by a one-step peptide-mediated biomineralization with the dye (TRAMA)-tagged peptides specific to caspase 3 as both template agents and the signal switch. Unlike conventional FRET-based fluorescent probes of caspase activity, these nanoprobes relied on the unique quenching effect of AuNCs through the nanosurface energy transfer (NSET) from dye to AuNCs. Intracellular caspase 3 activation cleaved the substrate peptide and released the dye from AuNCs, leading to a significant fluorescence lighting-up for sensitive and continuous analysis of caspase 3 activity in live cells, with a high signal-background ratio, wide linear range (32 pM–10 nM), and ultralow detection limit (12 pM). Moreover, this versatile AuNCs nanoprobe can serve as a theranostic platform via codisplaying pro-apoptotic and detecting peptides, which allows in situ activation and real-time monitoring of apoptosis in cancer cells. These results indicate that the AuNCs nanoprobe provides a smart molecular imaging and therapeutic agent targeted to cell apoptosis, which has great potential for apoptosis-related diagnosis and precision chemotherapy.
Co-reporter:Daiqi Li;Guoyan Lu;Chunyang Lei;Zhen Wang;Lijun Li
Science China Chemistry 2016 Volume 59( Issue 7) pp:809-815
Publication Date(Web):2016 July
DOI:10.1007/s11426-016-5564-5
DNA methylation, catalyzed by DNA methyltransferases (MTases), is a key component of genetic regulation, and DNA MTases have been regarded as potential targets in anticancer therapy. Herein, based on our previously developed DNA-mediated supercharged green fluorescent protein (ScGFP)/graphene oxide (GO) interaction, coupled with methylation-initiated template- free DNA polymerization, we propose a novel fluorescence assay strategy for sensitive detection of DNA MTase activity. A hairpin DNA with a methylation-sensitive site and an amino-modified 3'-terminal (DNA-1) was designed and worked as a starting molecule. In the presence of DNA MTase, methylation-sensitive restriction endonuclease, and terminal deoxynucleotidyl transferase (TdT), DNA-1 can be sequentially methylated, cleaved, and further elongated. The resulting long DNA fragments quickly bind with ScGFP and form the ScGFP/DNA nanocomplex. Such nanocomplex can effectively protect ScGFP from being adsorbed and quenched by GO. Without the methylation-initiated DNA polymerization, the fluorescence of ScGFP will be quenched by GO. Thus, the DNA MTase activity, which is proportional to the amount of DNA polymerization products, can be measured by reading the fluorescence of ScGFP/GO. The method was successfully used to detect the activity of DNA adenine methylation (Dam) MTase with a wide linear range (0.1–100 U/mL) and a low detection limit of 0.1 U/mL. In addition, the method showed high selectivity and the potential to be applied in a complex sample. Furthermore, this study was successfully extended to evaluate the inhibition effect of 5-fluorouracil on Dam MTase activity and detect TdT activity.
Co-reporter:Qin Wang, Hongjun Chen, Yong Li, Huixia Wang, Zhou Nie, Yufang Hu, Shouzhou Yao
Talanta 2016 Volume 161() pp:583-591
Publication Date(Web):1 December 2016
DOI:10.1016/j.talanta.2016.08.085
•An efficient electrochemical strategy on Citrate synthase was presented.•CoA is generated by CS-relevant catalysis reaction in TCA cycle.•The proposed platform is dependent on the formation of CoA-Ag(I) CP.•The developed sensor could be used for screening CS-related inhibitor.We report here a label-free and sensitive electrochemical method for probing Citrate synthase (CS) activity based on detailed investigations into the nucleic acid-mimicking coordination polymer (CP) formed from the coenzyme A (CoA)-Ag(I) repeat units. Our biosensing approach provides an especial and significant detection mechanism: CS can catalyze the essential condensation reaction between acetyl-coenzyme A (Ac-CoA) and oxaloacetate (OAA) to form citrate and CoA; then, in the presence of Ag(I), CoA-Ag(I) CP can be in situ formed because of the strong complexation ability of thiol groups of CoA toward Ag(I). The generated CoA-Ag(I) CP attaches to graphene-modified glassy carbon electrode surface by multiple adenine bases deriving from CoA and acting as the side groups along the polymeric backbone, which displays efficient H2O2-electrocatalyzing activity. More importantly, by using the formed polymer as signal output, the process is implemented to quantitatively analyze the activity of CS. Under the optimal conditions, CS with a detection limit as low as 0.00165 U/µL could be sensitively probed with a wide linear range from 0.0033 to 0.264 U/µL. Furthermore, with the character of label-free detection, high sensitivity and excellent selectivity, this strategy offers a convenient and specific method for CS activity detection and relevant inhibitors screening, which holds a promising potential in the practical application of CS-based biochemical research, disease diagnosis and drug discovery.
Co-reporter:Feipeng Jiao, Pin Qian, Yun Qin, Yalin Xia, Chunyan Deng, Zhou Nie
Talanta 2016 Volume 147() pp:98-102
Publication Date(Web):15 January 2016
DOI:10.1016/j.talanta.2015.09.045
•A novel biosensor for sensing uracil DNA glycosylase activity was developed.•The current responses of guanine bases in released ssDNA was as a signal indicator.•The electrochemical oxidation of guanine bases was greatly enhanced by graphene.•The label-free design did not need covalent attachment of redox indicator to DNA.•The UDG activity was simply determined with high sensitivity.Uracil-DNA glycosylase (UDG) as an important base excision repair enzymes is widely distributed in organism, and it plays a crucial role in sustaining the genome integrity. Therefore, it is significant to carry out the analysis of UDG activity. In this present work, a novel and label-free electrochemical sensing platform for the sensitive detection of uracil DNA glycosylase (UDG) activity has been developed. Herein, the graphene modified glassy carbon (GC) electrode was prepared. And two complementary DNA strands were hybridized to form dsDNA (P1P2). In the presence of UDG, the uracil bases in P1P2 were specifically hydrolyzed, inducing the unwinding of the DNA duplex, and accompanied by the release of P1. Thus, the released P1 was adsorbed onto the graphene/GC electrode surface via π–π stacking. By investigating the electrochemical behavior of P1 at the graphene/GC electrode, the electrochemical oxidation of guanine bases in P1 was obviously observed. Therefore, using the current responses of guanine base in P1 as a signal indicator, UDG activity can be simply determined with high sensitivity, and the detectable lowest concentration is 0.01 U/mL. This present design does not need covalent attachment of redox indicator to DNA, preventing participation of redox labels in the background. Meanwhile, the proposed strategy for the assay of UDG activity also has a remarkable sensitivity due to the excellent properties of graphene, which could increase both the immobilization amount of released ssDNA and the conductivity of the sensing system. All these elucidate that this developed protocol may lay a potential foundation for the sensitive detection of UDG activity in clinical diagnosis.Schematic representation of the electrochemical assay of UDG activity using the graphene modified GC electrode.
Co-reporter:Kaiyu He, Yong Li, Binbin Xiang, Peng Zhao, Yufang Hu, Yan Huang, Wang Li, Zhou Nie and Shouzhuo Yao
Chemical Science 2015 vol. 6(Issue 6) pp:3556-3564
Publication Date(Web):08 Apr 2015
DOI:10.1039/C5SC00371G
Molecular logic gates are capable of performing various logic tasks for biomarker detection, disease diagnostics and therapy, and controlling biological progress. Herein, we integrated multiple components of a logic device into a single DNA 3D nano-assembly with a triangular prism structure. Compared with the separate construction of each component in previously reported DNA logic gate systems, such an integrated design strategy made the 3D DNA nanoprism universal for logic gates, it can be reconfigured to execute diverse logic operations. Binary basic logic gates (OR, AND, INHIBIT and XOR), combinatorial gates (INHIBIT–OR), and multi-valued logic gates (ternary INHIBIT gate) were readily achieved by taking this DNA nanoprism as a universal platform. Moreover, a logic gate system for identification of even numbers and odd numbers from natural numbers was established successfully by employing only this single DNA nanoprism and four short single-stranded DNA. The universality of this nanoprism greatly simplified the design of DNA logic gate system. Additionally, this nanoprism was able to perform logic operation steadily in a biological matrix, indicating that this box-like DNA nanostructure applies to logic gates in a complicated environment. This study provided a unique opportunity to design versatile 3D DNA nanostructure-based intelligent nanodevices, which show great potential in biocomputing, multi-parameter sensing, and intelligent disease diagnostics and therapy.
Co-reporter:Yufang Hu, Siyu Chen, Yitao Han, Hongjun Chen, Qin Wang, Zhou Nie, Yan Huang and Shouzhuo Yao
Chemical Communications 2015 vol. 51(Issue 99) pp:17611-17614
Publication Date(Web):28 Sep 2015
DOI:10.1039/C5CC06593C
A nucleic acid-mimicking CoA–Ag(I) coordination polymer (CP) was in situ prepared and its unique electrocatalytic activity to H2O2 reduction was discovered. Based on it, a novel, label-free electrochemical sensor has been developed for the sensitive detection of coenzyme A (CoA) and histone acetyltransferase (HAT) activity.
Co-reporter:Shiyun Tang, Zhou Nie, Wang Li, Daiqi Li, Yan Huang and Shouzhuo Yao
Chemical Communications 2015 vol. 51(Issue 76) pp:14389-14392
Publication Date(Web):03 Aug 2015
DOI:10.1039/C5CC04170H
A label-free and convenient strategy for PARP-1 activity assay and inhibitors assessment has been developed based on the fluorescence resonance energy transfer (FRET) between a cationic conjugated polymer (CCP) and supercharged green fluorescent protein (scGFP).
Co-reporter:Zhen Wang, Yong Li, Lijun Li, Daiqi Li, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2015 vol. 51(Issue 69) pp:13373-13376
Publication Date(Web):14 Jul 2015
DOI:10.1039/C5CC04759E
The interaction between supercharged green fluorescent protein (ScGFP) and graphene oxide (GO) as well as the resulting quenching effect of GO on ScGFP were investigated. Based on this unique quenching effect and the DNA-mediated ScGFP/GO interaction, a label-free fluorescence method has been established for homogeneously assaying the activity and inhibition of base excision repair enzyme.
Co-reporter:Siyu Chen, Yong Li, Yufang Hu, Yitao Han, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2015 vol. 51(Issue 21) pp:4469-4472
Publication Date(Web):03 Feb 2015
DOI:10.1039/C5CC00067J
A novel and label-free fluorescence assay for histone acetyltransferase (HAT) activity was established via in situ generation of a nucleic acid-mimicking CoA–Au(I) coordination polymer (CP). Moreover, the potency of this assay for HAT-targeted drug discovery was proved by screening HAT inhibitors.
Co-reporter:Peng Zhao, Kaiyu He, Yitao Han, Zhen Zhang, Mengze Yu, Honghui Wang, Yan Huang, Zhou Nie, and Shouzhuo Yao
Analytical Chemistry 2015 Volume 87(Issue 19) pp:9998
Publication Date(Web):September 11, 2015
DOI:10.1021/acs.analchem.5b02614
Near-infrared (NIR) quantum dots (QDs) have emerged as an attractive bioimaging toolkit for exploring biological events because they can provide deep imaging penetration and low fluorescence background. However, the quantitation process of such NIR QDs generally relies on single-emission intensity change, which is susceptible to a variety of environmental factors. Herein, for the first time, we proposed a protein-directed co-template strategy to synthesize a NIR-based, dual-emission fluorescent nanohybrid (DEFN) constructed from far-red gold nanoclusters and NIR PbS QDs (AuNCs-PbS-QDs). The convenient protein-directed co-template synthesis avoids the tedious chemical coupling and modification required in conventional preparation approaches of DEFNs. Additionally, the dual-emission signals of AuNCs-PbS-QDs exhibit two well-resolved emission peaks (640 and 813 nm) separated by 173 nm, which can eliminate environmental interferences by the built-in correction of ratiometric signal, resulting in a more favorable system for bioimaging and biosensing. Next, the target-responsive capability of this NIR-based DEFN to ascorbic acid (AA) was discovered, enabling the proposed DEFN to ratiometrically detect AA with a linear range of 3–40 μM and a detection limit of 1.5 μM. This DEFN sensor possesses high selectivity, rapid response, and excellent photostability. Moreover, the feasibility of this NIR nanosensor has been fully proved by the ratiometric detection of AA for fruit internal quality assessment, in vitro cellular imaging, and in vivo imaging in nude mice.
Co-reporter:Yitao Han, Hao Li, Yufang Hu, Pei Li, Huixia Wang, Zhou Nie, and Shouzhuo Yao
Analytical Chemistry 2015 Volume 87(Issue 18) pp:9179
Publication Date(Web):August 26, 2015
DOI:10.1021/acs.analchem.5b01338
Protein acetylation of histone is an essential post-translational modification (PTM) mechanism in epigenetic gene regulation, and its status is reversibly controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Herein, we have developed a sensitive and label-free time-resolved luminescence (TRL) biosensor for continuous detection of enzymatic activity of HATs and HDACs, respectively, based on acetylation-mediated peptide/DNA interaction and Tb3+/DNA luminescent probes. Using guanine (G)-rich DNA-sensitized Tb3+ luminescence as the output signal, the polycationic substrate peptides interact with DNA with high affinity and subsequently replace Tb3+, eliminating the luminescent signal. HAT-catalyzed acetylation remarkably reduces the positive charge of the peptides and diminishes the peptide/DNA interaction, resulting in the signal on detection via recovery of DNA-sensitized Tb3+ luminescence. With this TRL sensor, HAT (p300) can be sensitively detected with a wide linear range from 0.2 to 100 nM and a low detection limit of 0.05 nM. The proposed sensor was further used to continuously monitor the HAT activity in real time. Additionally, the TRL biosensor was successfully applied to evaluating HAT inhibition by two specific inhibitors, anacardic acid and C464, and satisfactory Z′-factors above 0.73 were obtained. Moreover, this sensor is feasible to continuously monitor the HDAC (Sirt1)-catalyzed deacetylation with a linear range from 0.5 to 500 nM and a detection limit of 0.5 nM. The proposed sensor is a convenient, sensitive, and mix-and-read assay, presenting a promising platform for protein acetylation-targeted epigenetic research and drug discovery.
Co-reporter:Chao Yin, Ming Wang, Chunyang Lei, Zhen Wang, Pei Li, Yong Li, Wang Li, Yan Huang, Zhou Nie, and Shouzhuo Yao
Analytical Chemistry 2015 Volume 87(Issue 12) pp:6311
Publication Date(Web):May 11, 2015
DOI:10.1021/acs.analchem.5b01160
Protein phosphorylation catalyzed by protein kinases plays a critical role in many intracellular processes, and detecting kinase activity is important in biochemical research and drug discovery. Herein, we developed a novel fluorescent biosensor to detect protein kinase activity based on phosphorylation-mediated assembly of semisynthetic green fluorescent protein (GFP). A chimaera S-peptide composed of the 10th β-strand of GFP (s10) and a kinase substrate peptide was synthesized. Kinase-catalyzed phosphorylation of the S-peptide can protect its s10 part against cleavage by carboxypeptidase Y (CPY). Then, the peptide can bind the truncated GFP (tGFP, GFP without s10) to assemble intact GFP and recover fluorescence. Unphosphorylated S-peptide would be degraded by CPY, and fluorescent protein assembly could not occur. Thus, the kinase-catalyzed phosphorylation can switch on the fluorescence signal. This platform has been successfully applied to detect the activity of cAMP-dependent protein kinase with a low detection limit of 0.50 mU/μL and its inhibition of H-89 with an IC50 value of 23.4 nM. The feasibility of this method has been further demonstrated by assessment of the kinase activity and inhibition in the cell lysate. Moreover, based on the reverse principle, this method was expanded to detect the activity of protein phosphatase 1. Our method, using semisynthetic GFP as a readout, is facile, sensitive, label-free, and highly versatile, thus showing great potential as a promising platform for protein kinase detection and inhibitor screening.
Co-reporter:Yufang Hu, Qinpeng Shen, Wang Li, Zhuoliang Liu, Zhou Nie, Shouzhuo Yao
Biosensors and Bioelectronics 2015 Volume 63() pp:331-338
Publication Date(Web):15 January 2015
DOI:10.1016/j.bios.2014.07.066
•A novel electrochemical DNA sensor was developed based on TdT cascade amplification.•The dendritic DNA matrix with gold nanoparticles was used for signal amplification.•The sequence composition of TdT-yielded DNA is controlled by substrate dNTPs pool.•Crystal violet binds to TdT-synthesized G-rich sequence for signal readout.•This biosensing platform is versatile for detection of DNA, Pb2+, and thrombin.We describe a novel label-free amplified multifunctional strategy of dendritic electrochemical DNA sensor based on terminal deoxynucleotidyl transferase (TdT). We have found that the sequence composition of TdT-yielded DNA is largely dependent on the constitution of substrate deoxynucleotides (dNTPs) pool. After rational design of dNTPs pool and controllable TdT polymerization, dendritic protocol has been developed involving two-type amplification strategies; one is the formation of “trunk” and “branch” of the dendritic electrochemical sensor by TdT amplification; the other is the introduction of nucleic acid functionalized Au nanoparticles (DNA-AuNPs) for multiple branching. The results indicate that the G-rich ssDNA, which is synthesized under the condition of 40% deoxyadenosine triphosphate (dATP) and 60% deoxyguanosine triphosphate (dGTP), can be induced to form a long signal strand to G-quadruples (G4) in the presence of Pb2+. The electrochemical sensing platform is employed for sequence-specific DNA detection and the detection limit is as low as 1 fM. Our multifunctional strategy is further extended to Pb2+ detection and thrombin aptasensor. This proposed sensor displays excellent sensitivity and selectivity, and is applied for detection in complicated samples successfully.
Co-reporter:Zhuoliang Liu, Wang Li, Zhou Nie, Feifei Peng, Yan Huang and Shouzhuo Yao
Chemical Communications 2014 vol. 50(Issue 52) pp:6875-6878
Publication Date(Web):08 May 2014
DOI:10.1039/C4CC03103B
Randomly arrayed G-quadruplexes can serve as an efficient peroxidase-mimic DNAzyme and provide a novel and facile method to detect terminal deoxynucleotidyl transferase (TdT). Moreover, this G-rich sequence binding to the thioflavin T (ThT) dye can be applied in real-time fluorescent detection of TdT activity.
Co-reporter:Yanjuan Zhou, Kaiyu He, Shengquan Liu, Yong Li, Zhou Nie, Yan Huang and Shouzhuo Yao
RSC Advances 2014 vol. 4(Issue 36) pp:18668-18675
Publication Date(Web):08 Apr 2014
DOI:10.1039/C4RA01928H
Plasmonic changes in the growth process of gold nanoparticles (AuNPs) are not only intriguing for materials science, but also promising for developing new colorimetric biosensors and logic devices. Herein, we report a novel AuNP-based plasmonic phenomenon, namely chemical colorimetric square wave (CCSW) and further develop a series of label-free and simple colorimetric logic gates based on the CCSW. The CCSW refers to the unique phenomenon of silver ion concentration or pH-dependent discontinuous colorimetric signal change (blue-red-blue) observed in the hydrogen peroxide-mediated AuNP generation process. By setting blue and red colors as the OFF and ON colorimetric signals, respectively, the colorimetric signal profile of CCSW can be outputted in the format of OFF–ON–OFF, presenting an analog of an electronic square wave. Thereby, two CCSWs were successfully constructed, namely Ag+-CCSW and pH-CCSW. Subsequently, colorimetric AND, INHIBIT and OR logic gates were built via rational integration of two CCSWs. Unlike traditional AuNP-based logic gates with target-triggered AuNP crosslinking, these CCSW-based logic gates relied on the controllable growth of AuNPs, which are facile, fast and cost-efficient, avoiding tedious modification of AuNP probes and complicated operations. Therefore, our proposed CCSW and its derived logic gates present a promising application of plasmonic noble metal nanomaterials.
Co-reporter:Dr. Zhou Nie;Pengfei Wang;Cheng Tian;Dr. Chengde Mao
Angewandte Chemie International Edition 2014 Volume 53( Issue 32) pp:8402-8405
Publication Date(Web):
DOI:10.1002/anie.201404307
Abstract
Herein, we report a strategy for the synchronization of two self-assembly processes to assemble stimulus-responsive DNA nanostructures under isothermal conditions. We hypothesized that two independent assembly processes, when brought into proximity in space, could be synchronized and would exhibit positive synergy. To demonstrate this strategy, we assembled a ladderlike DNA nanostructure and a ringlike DNA nanostructure through two hybridization chain reactions (HCRs) and an HCR in combination with T-junction cohesion, respectively. Such proximity-induced synchronization adds a new element to the tool box of DNA nanotechnology. We believe that it will be a useful approach for the assembly of complex and responsive nanostructures.
Co-reporter:Chunyang Lei;Dr. Xiahong Xu;Jiang Zhou;Xin Liu;Dr. Zhou Nie;Meng Qing;Pei Li;Dr. Yan Huang; Shouzhuo Yao
Chemistry – An Asian Journal 2014 Volume 9( Issue 9) pp:2560-2567
Publication Date(Web):
DOI:10.1002/asia.201402221
Abstract
Protein kinase plays a vital role in regulating signal-transduction pathways and its simple and quick detection is highly desirable because traditional kinase assays typically rely on a time-consuming kinase-phosphorylation process (ca. 1 h). Herein, we report a new and rapid fluorescence-based sensing platform for probing the activity of protein kinase that is based on the super-quenching capacity of graphene oxide (GO) nanosheets and specific recognition of the aptameric peptide (FITC-IP20). On the GO/peptide platform, the fluorescence quenching of FITC-IP20 that is adsorbed onto GO can be restored by selective binding of active protein kinase to the aptameric peptide, thereby resulting in the fast switch-on detection of kinase activity (ca. 15 min). The feasibility of this method has been demonstrated by the sensitive measurement of the activity of cAMP-dependent protein kinase (PKA), with a detection limit of 0.053 mU μL−1. This assay technique was also successfully applied to the detection of kinase activation in cell lysate.
Co-reporter:Jiang Zhou, Xiahong Xu, Xin Liu, Hao Li, Zhou Nie, Meng Qing, Yan Huang, Shouzhuo Yao
Biosensors and Bioelectronics 2014 Volume 53() pp:295-300
Publication Date(Web):15 March 2014
DOI:10.1016/j.bios.2013.09.070
Co-reporter:Qinpeng Shen, Lifen Zhou, Yijia Yuan, Yan Huang, Binbin Xiang, Chunyan Chen, Zhou Nie, Shouzhuo Yao
Biosensors and Bioelectronics 2014 Volume 55() pp:187-194
Publication Date(Web):15 May 2014
DOI:10.1016/j.bios.2013.12.019
•A novel fluorescent copper ion biosensor relied on DNA-templated click reaction mediated intra-molecular G-quadruplex structure formation.•This “mix-and-read” signal-on detection is straightforward and homogenous based on the G-quadruplex-specific fluorogenic property of crystal violet.•High sensitivity due to the high yielding capability of DNA-templated organic synthesis (DTS).•Highly selective for copper ion because of the great specificity of click chemistry.A novel homogenous fluorescent sensor for signal-on detection of Cu2+ has been developed based on intra-molecular G-quadruplex formed by DNA-templated click reaction and crystal violet (CV) as label-free signal reporter. The clickable DNA probe consists of two G-rich strands (A and B) bearing azide and alkyne group, respectively, and a template strand (C) locating two proximate reactants by pairing with A and B. The sequences of A and B are derived from asymmetric split of the G-quadruplex sequence (TTAGGG)4. In the presence of Cu2+, the whole G-quadruplex sequence A–B is generated by chemical ligation of A and B via copper ion-catalyzed alkyne-azide cycloaddition, then released from template by toehold strand displacement, and consequently forming a stable intra-molecular G-quadruplex, which binds with CV to generate a strong fluorescent signal. Oppositely, weak fluorescence was obtained without Cu2+ because of unstable intermolecular G-quadruplex formed by A and B and lack of lateral loop connection. Therefore, the Cu2+ can be sensitively and specifically detected by the fluorescence of the CV-stained G-quadruplex with a low detection limit of 65 nM and a linear range of 0.1–3 µM. This method rationally integrated the DNA-templated synthesis and G-quadruplex structure-switch, presenting a simple and promising approach for biosensor development.
Co-reporter:Xin Liu, Yong Li, Xiahong Xu, Pei Li, Zhou Nie, Yan Huang, Shouzhuo Yao
TrAC Trends in Analytical Chemistry 2014 Volume 58() pp:40-53
Publication Date(Web):June 2014
DOI:10.1016/j.trac.2014.01.009
•Summarizing current applications of nanomaterials in protein kinase assays.•Classifying kinase nano-sensors according to analytical techniques.•Discussing the relative advantages and the limitations of these methods.•Predicting the trend in future development of nanomaterial-based kinase biosensors.Protein phosphorylation catalyzed by protein kinase is one of the most important post-translational modifications and plays a significant regulatory role in many vital biological processes. Aberrant protein-phosphorylation states and kinase activity are closely associated with many human diseases. Monitoring the kinase activity and its inhibition is essential for fundamental biochemical research and kinase-targeted drug discovery. Nanomaterial-based kinase assays provide a promising toolkit for exploring protein kinase functions, which have attracted growing interest in academic research, biomedical diagnosis, and pharmaceutical discovery. This article reviews the recent advances in the development of the protein kinase activity assays based on various nanomaterials, classifies these methods by different analytical techniques, summarizes the general design strategies, and offers perspectives on future developments.
Co-reporter:Yijia Yuan, Wenhua Li, Zhuoliang Liu, Zhou Nie, Yan Huang, Shouzhuo Yao
Biosensors and Bioelectronics 2014 Volume 61() pp:321-327
Publication Date(Web):15 November 2014
DOI:10.1016/j.bios.2014.05.038
•A novel synthesis strategy for silver nanoclusters based on enzymatically-generated DNA template was developed.•It is the first time to realize the turn-on detection of TdT and nuclease activity based on DNA–Ag NCs.•High sensitivity and selectivity is achieved in the detection application because of the turn-on design with high signal-to-background ratio.•The fluorescence intensity of DNA–AgNCs encapsulated by long-chained DNA was much stronger than those encapsulated by short-chained one.In the present day, oligonucleotide-encapsulated silver clusters (DNA–AgNCs) have been widely applied into bio-analysis as a signal producer. Herein, we developed a novel method to synthesize DNA–AgNCs encapsulated by long-chain cytosine (C)-rich DNA. Such DNA was polymerized in a template-free way by terminal deoxynucleotidyl transferase (TdT). We demonstrated that TdT-polymerized long chain C-rich DNA can serve as an excellent template for AgNCs synthesis. Based on this novel synthesis strategy, we developed a label-free and turn-on fluorescence assay to detect TdT activity with ultralow limit of detection (LOD) of 0.0318 U and ultrahigh signal to background (S/B) of 46.7. Furthermore, our proposed method was extended to a versatile biosensing strategy for turn-on nucleases activity assay based on the enzyme-activated TdT polymerization. Two nucleases, EcoRI and ExoIII as model of endonuclease and exonuclease, respectively, have been detected with high selectivity and competitive low LOD of 0.0629 U and 0.00867 U, respectively. Our work demonstrates the feasibility of TdT polymerization-based DNA–AgNCs synthesis strategy as a versatile and potent biosensing platform to detect the activity of DNA-related enzymes.
Co-reporter:Chunyang Lei;Dr. Yan Huang;Dr. Zhou Nie;Jun Hu;Lijun Li;Guoyan Lu;Yitao Han;Dr. Shouzhuo Yao
Angewandte Chemie International Edition 2014 Volume 53( Issue 32) pp:8358-8362
Publication Date(Web):
DOI:10.1002/anie.201403615
Abstract
Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio-macromolecules into cells. As a proof-of-principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluorescent protein (ScGFP) as the signal reporter, a simple turn-on homogenous method for DNA detection has been developed based on the polyionic nanoscale complex of ScGFP/DNA and toehold strand displacement. This assay shows high sensitivity and potent ability to detect single-base mismatch. Furthermore, combined with bisulfite conversion, this ScGFP-based assay was further applied to analyze site-specific DNA methylation status of genomic DNA extracted from real human colon carcinoma tissue sample with ultrahigh sensitivity (4 amol methylated DNA).
Co-reporter:Dr. Zhou Nie;Pengfei Wang;Cheng Tian;Dr. Chengde Mao
Angewandte Chemie 2014 Volume 126( Issue 32) pp:8542-8545
Publication Date(Web):
DOI:10.1002/ange.201404307
Abstract
Herein, we report a strategy for the synchronization of two self-assembly processes to assemble stimulus-responsive DNA nanostructures under isothermal conditions. We hypothesized that two independent assembly processes, when brought into proximity in space, could be synchronized and would exhibit positive synergy. To demonstrate this strategy, we assembled a ladderlike DNA nanostructure and a ringlike DNA nanostructure through two hybridization chain reactions (HCRs) and an HCR in combination with T-junction cohesion, respectively. Such proximity-induced synchronization adds a new element to the tool box of DNA nanotechnology. We believe that it will be a useful approach for the assembly of complex and responsive nanostructures.
Co-reporter:Chunyang Lei;Dr. Yan Huang;Dr. Zhou Nie;Jun Hu;Lijun Li;Guoyan Lu;Yitao Han;Dr. Shouzhuo Yao
Angewandte Chemie 2014 Volume 126( Issue 32) pp:8498-8502
Publication Date(Web):
DOI:10.1002/ange.201403615
Abstract
Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio-macromolecules into cells. As a proof-of-principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluorescent protein (ScGFP) as the signal reporter, a simple turn-on homogenous method for DNA detection has been developed based on the polyionic nanoscale complex of ScGFP/DNA and toehold strand displacement. This assay shows high sensitivity and potent ability to detect single-base mismatch. Furthermore, combined with bisulfite conversion, this ScGFP-based assay was further applied to analyze site-specific DNA methylation status of genomic DNA extracted from real human colon carcinoma tissue sample with ultrahigh sensitivity (4 amol methylated DNA).
Co-reporter:Fenfang Li, Chunyang Lei, Qinpeng Shen, Lijun Li, Ming Wang, Manli Guo, Yan Huang, Zhou Nie and Shouzhuo Yao
Nanoscale 2013 vol. 5(Issue 2) pp:653-662
Publication Date(Web):12 Nov 2012
DOI:10.1039/C2NR32156D
The rapid development in nanoparticle production and application during the past decade requires an easy, rapid, and predictive screening method for nanoparticles toxicity assay. In this study, the toxicological effects and the source of toxicity of copper nanoparticles (CuNPs) are investigated based on a stress-responsive bacterial biosensor array. According to the responses of the biosensing strains, it is found that CuNPs induce not only oxidative stress in E. coli, but also protein damage, DNA damage, and cell membrane damage, and ultimately cause cell growth inhibition. Through enzyme detoxification analysis, the toxicological effects of CuNPs are traced to H2O2 generation from CuNPs. Rapid copper release from CuNPs and Cu(I) production are observed. The oxidation of the released Cu(I) has a close relation to H2O2 production, as tris-(hydroxypropyltriazolylmethyl) amine, the specific Cu(I) chelator, can largely protect the cells from the toxicity of CuNPs. In addition, the TEM study shows that CuNPs can be adsorbed and incepted fast by the cells. Comparatively, copper microparticles are relatively stable in the system and practically non-toxic, which indicates the importance of toxic estimation of materials at the nanoscale. In addition, the Cu(II) ion can induce protein damage, membrane damage, and slight DNA damage only at a relatively high concentration. The current study reveals the preliminary mechanism of toxicity of CuNPs, and suggests that the stress-responsive bacterial biosensor array can be used as a simple and promising tool for rapid screening in vitro toxicity of nanoparticles and studying the primary mechanism of the toxicity.
Co-reporter:Xin Liu, Shuya Zhang, Penglong Tan, Jiang Zhou, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2013 vol. 49(Issue 18) pp:1856-1858
Publication Date(Web):14 Jan 2013
DOI:10.1039/C3CC38476D
A simple colorimetric assay for blood glucose has been developed based on enzymatic engineering of gold nanorods by glucose oxidase (GOx)-mediated oxidative etching of gold nanorods.
Co-reporter:Jiang Zhou, Xiahong Xu, Wei Liu, Xin Liu, Zhou Nie, Meng Qing, Lihua Nie, and Shouzhuo Yao
Analytical Chemistry 2013 Volume 85(Issue 12) pp:5746
Publication Date(Web):May 23, 2013
DOI:10.1021/ac400336u
The research on complicated kinomics and kinase-target drug discovery requires the development of simple, cost-effective, and multiplex kinase assays. Herein, we propose a novel and versatile biosensing platform for the detection of protein kinase activity based on graphene oxide (GO)–peptide nanocomplex and phosphorylation-induced suppression of carboxypeptidase Y (CPY) cleavage. Kinase-catalyzed phosphorylation protects the fluorophore-labeled peptide probe against CPY digestion and induces the formation of a GO/peptide nanocomplex resulting in fluorescence quenching, while the nonphosphopeptide is degraded by CPY to release free fluorophore as well as restore fluorescence. This GO-based nanosensor has been successfully applied to sensitively detect two model kinases, casein kinase (CKII) and cAMP–dependent protein kinase (PKA) with low detection limits of 0.0833 mU/μL and 0.134 mU/μL, respectively. The feasibility of this GO-based sensor was further demonstrated by the assessment of kinase inhibition by staurosporine and H-89, in vitro kinase assay in cell lysates, and simultaneous detection of CKII and PKA activity. Moreover, the GO-based fluorescence anisotropy (FA) kinase assay has been also developed using GO as a FA signal amplifier. The proposed sensor is homogeneous, facile, universal, label-free, and applicable for multiplexed kinase assay, presenting a promising method for kinase-related biochemical fundamental research and inhibitor screening.
Co-reporter:Peng Zhao, Lifen Zhou, Zhou Nie, Xiahong Xu, Wang Li, Yan Huang, Kaiyu He, and Shouzhuo Yao
Analytical Chemistry 2013 Volume 85(Issue 13) pp:6279
Publication Date(Web):June 6, 2013
DOI:10.1021/ac4004437
In this paper, the efficient quenching effect of deoxyguanosine-5′-phosphate (dGMP) on anodic electrochemiluminescence (ECL) of the CdTe/ZnS quantum dots (QDs) is reported for the first time. This ECL quenching was found to be specific for free dGMP and not observed for dGMP residues in different DNA structures. The unique dGMP-based QDs ECL quenching was then utilized to develop a versatile biosensing strategy to determine various protein–DNA interactions with the assistance of exonuclease, Exo I, to hydrolyze DNA and liberate dGMP. Taking single-stranded DNA binding protein (SSB) and thrombin as examples, two novel detection modes have been developed based on dGMP–QDs ECL strategy. The first method used hairpin probes and SSB-promoted probe cleavage by Exo I for facile signal-off detection of SSB, with a wide linear range of 1–200 nM and a low detection limit of 0.1 nM. The second method exploited aptamer–thrombin binding to protect probes against Exo I degradation for sensitive signal-on detection of thrombin, giving a linear response over a range of 1–150 nM and a detection limit as low as 0.1 nM. Both methods were homogeneous and label-free without QDs or DNA modification. Therefore, this dGMP-specific QDs ECL quenching presents a promising detection mechanism suitable for probing various protein–nucleic acid interactions.
Co-reporter:Yalin Xia, Wenhua Li, Ming Wang, Zhou Nie, Chunyan Deng, Shouzhuo Yao
Talanta 2013 Volume 107() pp:55-60
Publication Date(Web):30 March 2013
DOI:10.1016/j.talanta.2012.12.055
A novel, sensitive and enzymeless electrochemical sensor based on polynucleotide-templated silver nanoclusters (DNA-AgNCs)/graphene composite film was developed for the detection of hydrogen peroxide. The graphene modified glassy carbon electrode (GCE) was employed because graphene has several advantages including excellent conductivity, biocompatibility, and large surface area to volume ratio. In addition, it was found that DNA-AgNCs have remarkable electrocatalytic activity toward the reduction of hydrogen peroxide, and can be easily immobilized onto the surface of the graphene/GCE by π–π stacking. The sensor based on the (DNA-AgNCs)/graphene/GCE exhibited a rapid response (ca. 3 s), a low detection limit (3 μM), a wide linear range from 15 μM to 23 mM, high selectivity, as well as good repeatability. Moreover, the common interfering species, such as ascorbic acid, uric acid, dopamine, glutathione, and l-cysteine, did not result in any interference. This present work may expand the use of silver nanoclusters in the field of electrochemical sensor.Highlights► We fabricated the electrochemical sensor with simplicity and high sensitivity. ► DNA-AgNCs were simply synthesized and assembled onto graphene by π–π stacking. ► DNA-AgNCs possess enhanced electrocatalysis toward the reduction of H2O2. Graphene was electrodeposited onto the glassy carbon electrode. ► Graphene is useful for the immobilization of more DNA-AgNCs onto the electrode.
Co-reporter:Wenhua Li, Wang Li, Yufang Hu, Yalin Xia, Qinpeng Shen, Zhou Nie, Yan Huang, Shouzhuo Yao
Biosensors and Bioelectronics 2013 Volume 47() pp:345-349
Publication Date(Web):15 September 2013
DOI:10.1016/j.bios.2013.03.038
A novel label-free, rapid, cost-effective, and highly sensitive fluorometric sensor has been constructed for the detection of acetylcholinesterase (AChE) activity and its inhibitor based on the fluorescence quenching of DNA-templated copper/silver nanoclusters (DNA-Cu/AgNCs). In this assay, AChE catalyzes the hydrolysis of acetylthiocholine (ATCh) to form thiocholine which induces fluorescence quenching of DNA-Cu/AgNCs. The AChE activity could be detected as low as 0.05 mU/mL and with a linear range from 0.05 to 2.0 mU/mL. This assay offers a very convenient “mix and detect” approach for AChE activity. On the other hand, tacrine and organophosphorus pesticides (OPPs) were employed to inhibit the hydrolysis of ATCh, which could eliminate the fluorescence quenching of DNA-Cu/AgNCs. The IC50 of tacrine and methamidophos were estimated to be 16.9 nM and 0.075 mg/L, respectively. This method was also used to detect spiked OPPs in agricultural products successfully. The present work may expand the use of DNA-Cu/AgNCs to the field of enzyme sensors.
Co-reporter:Qinpeng Shen, Wenhua Li, Shiyun Tang, Yufang Hu, Zhou Nie, Yan Huang, Shouzhuo Yao
Biosensors and Bioelectronics 2013 Volume 41() pp:663-668
Publication Date(Web):15 March 2013
DOI:10.1016/j.bios.2012.09.032
A novel colorimetric copper(II) biosensor has been developed based on the high specificity of alkyne–azide click reaction to the catalysis of copper ions and unmodified gold nanoparticles (AuNPs) as the signal reporter. The clickable DNA probe consists of two parts: an azide group-modified double-stranded DNA (dsDNA) hybrid with an elongated tail and a short alkyne-modified single-stranded DNA (ssDNA). Because of low melting temperature of the short ssDNA, these two parts are separated in the absence of Cu2+. Copper ion-induced azide–alkyne click ligation caused a structural change of probe from the separated form to entire dsDNA form. This structural change of probe can be monitored by the unmodified AuNPs via mediating their aggregation with a red-to-blue colorimetric read-out because of the differential ability of ssDNA and dsDNA to protect AuNPs against salt-induced aggregation. Under the optimum conditions, this biosensor can sensitively and specifically detect Cu2+ with a low detection limit of 250 nM and a linear range of 0.5–10 μM. The method is simple and economic without dual-labeling DNA and AuNPs modification. It is also highly selective for Cu2+ in the presence of high concentrations of other environmentally relevant metal ions because of the great specificity of the copper-caused alkyne–azide click reaction, which potentially meets the requirement of the detection in real samples.
Co-reporter:Qinpeng Shen, Shiyun Tang, Wenhua Li, Zhou Nie, Zhuoliang Liu, Yan Huang and Shouzhuo Yao
Chemical Communications 2012 vol. 48(Issue 2) pp:281-283
Publication Date(Web):22 Nov 2011
DOI:10.1039/C1CC16049D
A novel fluorescent strategy has been developed for sensitive turn-on detection of Cu2+ based on high efficiency of DNA-templated organic synthesis, great specificity of alkyne–azide click reaction to the catalysis of copper ions and the sequential strand displacement for signal transduction.
Co-reporter:Xiahong Xu, Jiang Zhou, Xin Liu, Zhou Nie, Meng Qing, Manli Guo, and Shouzhuo Yao
Analytical Chemistry 2012 Volume 84(Issue 11) pp:4746
Publication Date(Web):April 26, 2012
DOI:10.1021/ac3001918
Protein kinases are significant regulators in the cell signal pathway, and it is difficult to achieve quick kinase detection because traditional kinase assays normally rely on a time-consuming kinase phosphorylation process. Herein, we present a novel one-step strategy to detect protein kinase by using a kinase-specific aptameric peptide-functionalized quartz crystal microbalance (QCM) electrode, in which the detection can be finished in less than 10 min. A peptide kinase inhibitor (IP20) was used as the aptameric peptide because of its selective and strong interaction with the target protein kinase (cyclic adenosine monophosphate-dependent protein kinase A, PKA), high stability, and ease of inexpensive synthesis, presenting a new direct recognition element for kinase. The aptameric peptide was immobilized on the Au-coated quartz electrode through dual-thiol anchoring and the binding of His-tagged peptide with a nitrilotriacetic acid/Ni(II) complex, fabricating a highly specific and stable detection platform. The interaction of aptameric peptide with kinase was monitored with the QCM in real time, and the concentration of protein kinase was sensitively measured by the frequency response of the QCM with the low detection limit for PKA at 0.061 mU μL–1 and a linear range from 0.64 to 22.33 mU μL–1. This method is rapid and reagentless and does not require a phosphorylation process. The versatility of our aptameric peptide-based strategy has also been demonstrated by the application in kinase assay using electrochemical impedance spectroscopy. Moreover, this method was successfully applied to detect the forskolin/3-isobutyl-1-methylxanthine-stimulated activation of PKA in cell lysate.
Co-reporter:Kaiyu He;Wang Li; Zhou Nie;Yan Huang;Zhuoliang Liu;Lihua Nie ; Shouzhuo Yao
Chemistry - A European Journal 2012 Volume 18( Issue 13) pp:3992-3999
Publication Date(Web):
DOI:10.1002/chem.201102290
Abstract
The DNA nick repair catalyzed by DNA ligase is significant for fundamental life processes, such as the replication, repair, and recombination of nucleic acids. Here, we have employed ligase to regulate DNAzyme activity and developed a homogeneous, colorimetric, label-free and DNAzyme-based strategy to detect DNA ligase activity. This novel strategy relies on the ligation-trigged activation or production of horseradish peroxidase mimicking DNAzyme that catalyzes the generation of a color change signal; this results in a colorimetric assay of DNA ligase activity. Using T4 DNA ligase as a model, we have proposed two approaches to demonstrate the validity of the DNAzyme strategy. The first approach utilizes an allosteric hairpin-DNAzyme probe specifically responsive to DNA ligation; this approach has a wide detection range from 0.2 to 40 U mL−1 and a detection limit of 0.2 U mL−1. Furthermore, the approach was adapted to probe nucleic acid phosphorylation and single nucleotide mismatch. The second approach employs a “split DNA machine” to produce numerous DNAzymes after being reassembled by DNA ligase; this greatly enhances the detection sensitivity by a signal amplification cascade to achieve a detection limit of 0.01 U mL−1.
Co-reporter:Chunyan Deng, Yalin Xia, Chunhui Xiao, Zhou Nie, Minghui Yang, Shihui Si
Biosensors and Bioelectronics 2012 Volume 31(Issue 1) pp:469-474
Publication Date(Web):15 January 2012
DOI:10.1016/j.bios.2011.11.018
Based on the excellent physicochemical properties of boron-doped carbon nanotubes (BCNTs), the electrochemical analysis of four free DNA bases at the BCNTs modified glassy carbon (GC) electrode was investigated. Herein, the BCNTs/GC electrode exhibited remarkable electrocatalytic activity towards the oxidation of purine bases (guanine (G), adenine (A)). More significantly, the direct oxidation of pyrimidine bases (thymine (T), cytosine (C)) was realized. It may be due to that BCNTs have the advantages of high electron transfer kinetics, large surface area, prominent antifouling ability and electrode activity. On basis of this, a novel and simple strategy for the determination of G, A, T and C was proposed. The BCNTs/GC electrode showed high sensitivity, wide linear range and capability of detection for the electrochemical determination of G, A, T, and C. On the other hand, the electrochemical oxidation of quaternary mixture of G, A, T, and C at the BCNTs/GC electrode was investigated. It was obtained that the peak separation between G and A, A and T, T and C were large enough for their potential recognition in mixture without any separation or pretreatment. The BCNTs/GC electrode also displayed good stability, reproducibility and excellent anti-interferent ability. Therefore, it can be believed that the BCNTs/GC electrode would provide a potential application for the electrochemical detection of DNA in the field of genetic-disease diagnosis.
Co-reporter:Wang Li, Zhou Nie, Kaiyu He, Xiahong Xu, Yong Li, Yan Huang and Shouzhuo Yao
Chemical Communications 2011 vol. 47(Issue 15) pp:4412-4414
Publication Date(Web):10 Mar 2011
DOI:10.1039/C0CC05727D
A simple strategy based on unmodified AuNPs and specific Zn2+ binding peptide is exploited here as a new label-free mechanism for the rapid colorimetric Zn2+ assay.
Co-reporter:Xiahong Xu, Xin Liu, Zhou Nie, Yuliang Pan, Manli Guo, and Shouzhuo Yao
Analytical Chemistry 2011 Volume 83(Issue 1) pp:52
Publication Date(Web):December 3, 2010
DOI:10.1021/ac102786c
Herein, we present a novel label-free fluorescent assay for monitoring the activity and inhibition of protein kinases based on the aggregation behavior of unmodified CdTe quantum dots (QDs). In this assay, cationic substrate peptides induce the selective aggregation of unmodified QDs with anionic surface charge, whereas phosphorylated peptides do not. Phosphorylation by kinase alters the net charge of peptides and subsequently inhibits the aggregation of unmodified QDs, causing an enhanced fluorescence with a 45 nm blue-shift in emission and a yellow-to-green emission color change. Hence the fluorescence response allows this QD-based method to easily probe kinase activity by a spectrometer or even by the naked eye. The feasibility of the method has been demonstrated by sensitive measurement of the activity of cAMP-dependent protein kinase (PKA) with a low detection limit (0.47 mU μL−1). On the basis of the fluorescence response of QDs on the concentration of PKA inhibitor H-89, the IC50 value, the half maximal inhibitory concentration, was estimated, which was in agreement with the literature value. Moreover, the system can be applicable to detect the Forskolin/3-isobutyl-1-methylxantine (IBMX)-stimulated activation of PKA in cell lysate. Unlike the existing QD-based enzyme activity assays in which the modification process of QDs is essential, this method relies on unmodified QDs without the requirement of peptide labeling and QDs’ modification, presenting a promising candidate for cost-effective kinase activity and inhibitor screening assays.
Co-reporter:MengDong Wang;YiTao Han;XingXing Liu;ChunYan Deng
Science China Chemistry 2011 Volume 54( Issue 8) pp:
Publication Date(Web):2011 August
DOI:10.1007/s11426-011-4345-4
Based on the layer-by-layer self-assembly of positively charged cetyltrimethylammonium bromide (CTAB) wrapped gold nanorods (AuNRs) and negatively charged superoxide dismutase (SOD) from their aqueous solutions on cysteine modified gold electrode (Cys/Au), a third generation electrochemical biosensor ((SOD/AuNRs)2/Cys/Au) for superoxide anion (O2·−) was developed. The two layers assembly of SOD/AuNRs can significantly enhance the direct electron transfer between SOD and the electrode. The functional enzymatic activities of the SOD offer an electrochemical approach to the determination of O2·−. In the reductive regions, the proposed sensor exhibits excellent analytical performances, such as wide linear range (200 nM to 0.2 mM O2·−), low detection limit (100 nM O2·−), high sensitivity (22.11 nA cm−2 μM−1), short response time (less than 5 s), good stability and reproducibility, while no obvious interferences are caused by commonly met interfering species including hydrogen peroxide (H2O2), uric acid (UA) and ascorbic acid (AA).
Co-reporter:Xingxing Liu, Wang Li, Qinpeng Shen, Zhou Nie, Manli Guo, Yitao Han, Wei Liu, Shouzhuo Yao
Talanta 2011 Volume 85(Issue 3) pp:1603-1608
Publication Date(Web):15 September 2011
DOI:10.1016/j.talanta.2011.06.061
The heavy metal ions–nucleobases interaction is an important research topic in environmental and biochemical analysis. The presence of the silver ion (Ag+) may influence the formation of oxidation intermediate and the electrocatalytic oxidation activity of guanine (G), since Ag+ can interact with guanine at the binding sites which are involved in the electrocatalytic oxidation reaction of guanine. According to this principle, a new electrochemical sensor for indirectly detecting Ag+ based on the interaction of Ag+ with isolated guanine base using differential pulse voltammetry (DPV) was constructed. Among the heavy metal ions examined, only Ag+ showed the strongest inhibitory effect on the electrocatalytic oxidation of guanine at the multi-walled carbon nanotubes modified glassy carbon electrode (CNTs/GC). And the quantitative study of Ag+ based on Ag+–G sensing system gave a linear range from 100 nM to 2.5 μM with a detection limit of 30 nM. In addition, this modified electrode had very good reproducibility and stability. The developed electrochemical method is an ideal tool for Ag+ detection with some merits including remarkable simplicity, low-cost, and no requirement for probe preparation.
Co-reporter:Wang Li, Zhuoliang Liu, Hui Lin, Zhou Nie, Jinhua Chen, Xiahong Xu and Shouzhuo Yao
Analytical Chemistry 2010 Volume 82(Issue 5) pp:1935
Publication Date(Web):February 11, 2010
DOI:10.1021/ac902670c
DNA methylation catalyzed by methyltransferase (MTase) is a significant epigenetic process for modulating gene expression. Traditional methods to study MTase activity require a laborious and costly DNA labeling process. In this article, we report a simple, colorimetric, and label-free methylation-responsive DNAzyme (MR-DNAzyme) strategy for MTase activity analysis. This new strategy relies on horseradish peroxidase (HRP) mimicking DNAzyme and the methylation-responsive sequence (MRS) of DNA which can be methylated and cleaved by the MTase/endonuclease coupling reaction. Methylation-induced scission of MRS would activate the DNAzyme that can catalyze the generation of a color signal for the amplified detection of methylation events. Taking Dam MTase and DpnI endonuclease as examples, we have developed two colorimetric methods based on the MR-DNAzyme strategy. The first method is to utilize an engineered hairpin-DNAzyme hybrid probe for facile turn-on detection of Dam MTase activity, with a wide linear range (6−100 U/mL) and a low detection limit (6 U/mL). Furthermore, this method could be easily expanded to profile the activity and inhibition of restriction endonuclease. The second method involves a methylation-triggered DNAzyme-based DNA machine, which achieves the ultrahigh sensitive detection of Dam MTase activity (detection limit = 0.25 U/mL) by a two-step signal amplification cascade.
Co-reporter:Mengdong Wang, Yitao Han, Zhou Nie, Chunyang Lei, Yan Huang, Manli Guo, Shouzhuo Yao
Biosensors and Bioelectronics 2010 Volume 26(Issue 2) pp:523-529
Publication Date(Web):15 October 2010
DOI:10.1016/j.bios.2010.07.058
Study on antioxidants’ radical scavenging processes and antioxidant capabilities is important for understanding the protective role of antioxidants against oxidative damages associated with some chronic diseases and food degradation. Traditional methods to monitor the radical scavenging by antioxidant require expensive instrument and sophisticated synthesis process. Herein, we report a novel, simple, colorimetric DNAzyme-based method to detect radical-scavenging capacity of antioxidant. In this new strategy, horseradish peroxidase (HRP) mimicking DNAzyme catalyzes the oxidation of ABTS2− (2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)) by H2O2 to generate blue/green ABTS− radical, which can be scavenged by antioxidants resulting in color change. The typical kinetic curve of antioxidant-inhibited generation of ABTS− shows distinct biphasic pattern, involving a lag phase (stage I) and a linear increase phase (stage II). kt value, the product of lag time (t) and the slope of the curve in stage II (k), was used as the parameter for antioxidant capacity determination. This DNAzyme-based antioxidant assay has been effectively used to quantitatively detect the concentrations of antioxidants and evaluate the antioxidant capabilities of a variety of antioxidants and some real samples. Compared with traditional antioxidant assays, this method is thermostable, pH stable, and time-saving, which presents a promising platform for antioxidant assay.
Co-reporter:Qinpeng Shen, Zhou Nie, Manli Guo, Chuan-Jian Zhong, Bin Lin, Wang Li and Shouzhuo Yao
Chemical Communications 2009 (Issue 8) pp:929-931
Publication Date(Web):14 Jan 2009
DOI:10.1039/B818081D
The sequence-length-dependent adsorption of single-stranded DNA (ssDNA) on unmodified gold nanoparticles is exploited as a new mechanism for colorimetric nuclease assay and measurement of oxidative DNA damage.
Co-reporter:Xiahong Xu, Zhou Nie, Jinhua Chen, Yingchun Fu, Wang Li, Qinpeng Shen and Shouzhuo Yao
Chemical Communications 2009 (Issue 45) pp:6946-6948
Publication Date(Web):16 Oct 2009
DOI:10.1039/B913943E
A novel label-free electrochemical strategy for monitoring the activity and inhibition of protein kinase is developed, based on the linkage between the phosphorylated peptide and DNA functionalized Au nanoparticles (DNA–AuNPs) by Zr4+ and the chronocoulometric response of [Ru(NH3)6]3+ absorbed on the DNA–AuNPs.
Co-reporter:Chunyan Deng, Jinhua Chen, Lihua Nie, Zhou Nie and Shouzhuo Yao
Analytical Chemistry 2009 Volume 81(Issue 24) pp:9972
Publication Date(Web):November 19, 2009
DOI:10.1021/ac901727z
In this paper, a bifunctional electrochemical biosensor for highly sensitive detection of small molecule (adenosine) or protein (lysozyme) was developed. Two aptamer units for adenosine and lysozyme were immobilized on the gold electrode by the formation of DNA/DNA duplex. The detection of adenosine or lysozyme could be carried out by virtue of switching structures of aptamers from DNA/DNA duplex to DNA/target complex. The change of the interfacial feature of the electrode was characterized by cyclic voltammertic (CV) response of surface-bound [Ru(NH3)6]3+. On the other hand, DNA functionalized Au nanoparticles (DNA-AuNPs) were used to enhance the sensitivity of the aptasensor because DNA-AuNPs modified interface could load more [Ru(NH3)6]3+ cations. Thus, the assembly of two aptamer-contained DNA strands integrated with the DNA−AuNPs amplification not only improves the sensitivity of the electrochemical aptasensor but also presents a simple and general model for bifunctional aptasensor. The proposed aptasensor has low detection limit (0.02 nM for adenosine and 0.01 μg mL−1 for lysozyme) and exhibits several advantages such as high sensitivity and bifunctional recognition.
Co-reporter:Wang Li, Zhou Nie, Xiahong Xu, Qinpeng Shen, Chunyan Deng, Jinhua Chen, Shouzhuo Yao
Talanta 2009 Volume 78(Issue 3) pp:954-958
Publication Date(Web):15 May 2009
DOI:10.1016/j.talanta.2009.01.009
A sensitive, label free electrochemical aptasensor for small molecular detection has been developed in this work based on gold nanoparticles (AuNPs) amplification. This aptasensor was fabricated as a tertiary hybrid DNA–AuNPs system, which involved the anchored DNA (ADNA) immobilized on gold electrode, reporter DNA (RDNA) tethered with AuNPs and target-responsive DNA (TRDNA) linking ADNA and RDNA. Electrochemical signal is derived from chronocoulometric interrogation of [Ru(NH3)6]3+ (RuHex) that quantitatively binds to surface-confined DNA via electrostatic interaction. Using adenosine triphosphate (ATP) as a model analyte and ATP-binding aptamer as a model molecular reorganization element, the introduction of ATP triggers the structure switching of the TRDNA to form aptamer–ATP complex, which results in the dissociation of the RDNA capped AuNPs (RDNA–AuNPs) and release of abundant RuHex molecules trapped by RDNA–AuNPs. The incorporation of AuNPs in this strategy significantly enhances the sensitivity because of the amplification of electrochemical signal by the RDNA–AuNPs/RuHex system. Under optimized conditions, a wide linear dynamic range of 4 orders of magnitude (1 nM–10 μM) was reached with the minimum detectable concentration at sub-nanomolar level (0.2 nM). Those results demonstrate that our nanoparticles-based amplification strategy is feasible for ATP assay and presents a potential universal method for other small molecular aptasensors.
Co-reporter:MengDong Wang;ChunYan Deng;XiaHong Xu
Science China Chemistry 2009 Volume 52( Issue 11) pp:
Publication Date(Web):2009 November
DOI:10.1007/s11426-009-0250-5
Amino acid ionic liquids (AAILs) have attracted much attention due to their special chemical and physical properties, especially their outstanding biocompatibility and truly green aspect. In this work, a novel electrochemical biosensing platform based on AAILs/carbon nanotubes (CNTs) composite was fabricated. AAILs were used as a novel solvent for glucose oxidase (GOD) and the GOD-AAILs/CNTs/GC electrode was conveniently prepared by immersing the carbon nanotubes (CNTs) modified glassy carbon (GC) electrode into AAILs containing GOD. The direct electrochemistry of GOD on the GOD-AAILs/CNTs/GC electrode has been investigated and a pair of reversible peaks was obtained by cyclic voltammetry. The immobilized glucose oxidase could retain bioactivity and catalyze the reduction of dissolved oxygen. Due to the synergic effect of AAILs and CNTs, the GOD-AAILs/CNTs/GC electrode shows excellent electrocatalytic activity towards glucose with a linear range from 0.05 to 0.8 mM and a detection limit of 5.5 µM (S/N = 3). Furthermore, the biosensor exhibits good stability and ability to exclude the interference of commonly coexisting uric and ascorbic acid. Therefore, AAILs/CNTs composite can be a good candidate biocompatible material for the direct electrochemistry of the redox-active enzyme and the construction of third- generation enzyme sensors.
Co-reporter:Huixia Wang, Yong Li, Kunli Zhao, Siyi Chen, Qin Wang, Bin Lin, Zhou Nie, Shouzhuo Yao
Biosensors and Bioelectronics (15 May 2017) Volume 91() pp:
Publication Date(Web):15 May 2017
DOI:10.1016/j.bios.2016.12.065
•A fluorometric biosensor for HAT and HDAC detection was developed.•The mechanism is based on HATs/HDACs-mediated G4/ThT fluorescent switch.•This G4-based biosensor robust, homogeneous, facile, and cost-efficient.•A high-throughput screening of HAT/HDAC-targeted inhibitors was achieved.Reversible protein acetylation, one of the key types of post-translational modifications, is composed of histone acetylation and deacetylation, which is typically catalyzed by histone acetyltransferases (HATs) and histone deacetylases (HDACs) respectively. Herein, a label-free fluorescent method has been established for the homogeneous bioassay of HAT/HDAC activity and respective inhibitors. The proposed approach is primarily based on the electrostatic interaction between G-quadruplexes (G4s) and acetylation-related peptides, which results in marked change of fluorescent intensity of G4/Thioflavin T (ThT) complexes. This HAT (p300) activity assay is exceedingly sensitive and selective, with a linear range from 0.1 to 120 nM and a detection limit of 0.05 nM. Moreover, this biosensor is feasible to detect the HDAC (Sirt1) activity with a linear range from 1 to 450 nM and a detection limit of 1 nM. The potency of this assay is further demonstrated by detecting HAT/HDAC activity in cell lysates and evaluating HAT and HDAC-targeted inhibitors, C464 and EX 527, respectively. The proposed assay is convenient, label-free and cost-efficient, which is promising for HAT/HDAC-targeted epigenetic research and pharmaceutical development.
Co-reporter:Lifen Zhou, Qinpeng Shen, Peng Zhao, Bingbing Xiang, Zhou Nie, Yan Huang, Shouzhuo Yao
Methods (15 December 2013) Volume 64(Issue 3) pp:299-304
Publication Date(Web):15 December 2013
DOI:10.1016/j.ymeth.2013.09.001
•Cu2+ assay based on DNA templated click chemistry and graphene oxide nanosheet.•Highly selective for Cu2+ due to its catalyst specificity to click reaction.•Quick detection because of the accelerated reaction by DNA-templated synthesis.•High sensitivity resulted from the super-quenching ability of graphene oxide.A novel DNA-templated click chemistry strategy for homogenous fluorescent detection of Cu2+ has been developed based on click ligation-dependent DNA structure switch and the selective quenching ability of graphene oxide (GO) nanosheet. The clickable duplex probe consists of two DNA strands with alkyne and azide group, respectively, and Cu+-catalyzed alkyne–azide cycloaddition (CuAAC) reaction can chemically ligate these two strands. Toehold sequence displacement was consequently exploited to achieve DNA structure transformation bearing fluorescent tag FAM. Cu2+-induced chemical ligation caused the probe transfer to hybrid structure with single stranded DNA (ssDNA) tail, while only duplex structure was obtained without Cu2+. This structural difference can be probed by GO-based fluorescence detection due to the preferential binding of GO to ssDNA. Under the optimum conditions, this sensor can sensitively and specifically detect Cu2+ with a low detection limit of 58 nM and a linear range of 0.1–10 μM. This new strategy is highly sensitive and selective for Cu2+ detection because of the great specificity of click chemistry and super-quenching ability of GO. Moreover, with the aid of high efficient DNA templated synthesis, the detection process requires only about half an hour which is much quicker than previous click-chemistry-based Cu2+ sensors.Download full-size image
Co-reporter:Xin Liu, Shuya Zhang, Penglong Tan, Jiang Zhou, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2013 - vol. 49(Issue 18) pp:NaN1858-1858
Publication Date(Web):2013/01/14
DOI:10.1039/C3CC38476D
A simple colorimetric assay for blood glucose has been developed based on enzymatic engineering of gold nanorods by glucose oxidase (GOx)-mediated oxidative etching of gold nanorods.
Co-reporter:Siyu Chen, Yong Li, Yufang Hu, Yitao Han, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2015 - vol. 51(Issue 21) pp:NaN4472-4472
Publication Date(Web):2015/02/03
DOI:10.1039/C5CC00067J
A novel and label-free fluorescence assay for histone acetyltransferase (HAT) activity was established via in situ generation of a nucleic acid-mimicking CoA–Au(I) coordination polymer (CP). Moreover, the potency of this assay for HAT-targeted drug discovery was proved by screening HAT inhibitors.
Co-reporter:Zhen Wang, Yong Li, Lijun Li, Daiqi Li, Yan Huang, Zhou Nie and Shouzhuo Yao
Chemical Communications 2015 - vol. 51(Issue 69) pp:NaN13376-13376
Publication Date(Web):2015/07/14
DOI:10.1039/C5CC04759E
The interaction between supercharged green fluorescent protein (ScGFP) and graphene oxide (GO) as well as the resulting quenching effect of GO on ScGFP were investigated. Based on this unique quenching effect and the DNA-mediated ScGFP/GO interaction, a label-free fluorescence method has been established for homogeneously assaying the activity and inhibition of base excision repair enzyme.
Co-reporter:Yufang Hu, Siyu Chen, Yitao Han, Hongjun Chen, Qin Wang, Zhou Nie, Yan Huang and Shouzhuo Yao
Chemical Communications 2015 - vol. 51(Issue 99) pp:NaN17614-17614
Publication Date(Web):2015/09/28
DOI:10.1039/C5CC06593C
A nucleic acid-mimicking CoA–Ag(I) coordination polymer (CP) was in situ prepared and its unique electrocatalytic activity to H2O2 reduction was discovered. Based on it, a novel, label-free electrochemical sensor has been developed for the sensitive detection of coenzyme A (CoA) and histone acetyltransferase (HAT) activity.
Co-reporter:Zhuoliang Liu, Wang Li, Zhou Nie, Feifei Peng, Yan Huang and Shouzhuo Yao
Chemical Communications 2014 - vol. 50(Issue 52) pp:NaN6878-6878
Publication Date(Web):2014/05/08
DOI:10.1039/C4CC03103B
Randomly arrayed G-quadruplexes can serve as an efficient peroxidase-mimic DNAzyme and provide a novel and facile method to detect terminal deoxynucleotidyl transferase (TdT). Moreover, this G-rich sequence binding to the thioflavin T (ThT) dye can be applied in real-time fluorescent detection of TdT activity.
Co-reporter:Qinpeng Shen, Shiyun Tang, Wenhua Li, Zhou Nie, Zhuoliang Liu, Yan Huang and Shouzhuo Yao
Chemical Communications 2012 - vol. 48(Issue 2) pp:NaN283-283
Publication Date(Web):2011/11/22
DOI:10.1039/C1CC16049D
A novel fluorescent strategy has been developed for sensitive turn-on detection of Cu2+ based on high efficiency of DNA-templated organic synthesis, great specificity of alkyne–azide click reaction to the catalysis of copper ions and the sequential strand displacement for signal transduction.
Co-reporter:Qinpeng Shen, Zhou Nie, Manli Guo, Chuan-Jian Zhong, Bin Lin, Wang Li and Shouzhuo Yao
Chemical Communications 2009(Issue 8) pp:NaN931-931
Publication Date(Web):2009/01/14
DOI:10.1039/B818081D
The sequence-length-dependent adsorption of single-stranded DNA (ssDNA) on unmodified gold nanoparticles is exploited as a new mechanism for colorimetric nuclease assay and measurement of oxidative DNA damage.
Co-reporter:Wang Li, Zhou Nie, Kaiyu He, Xiahong Xu, Yong Li, Yan Huang and Shouzhuo Yao
Chemical Communications 2011 - vol. 47(Issue 15) pp:NaN4414-4414
Publication Date(Web):2011/03/10
DOI:10.1039/C0CC05727D
A simple strategy based on unmodified AuNPs and specific Zn2+ binding peptide is exploited here as a new label-free mechanism for the rapid colorimetric Zn2+ assay.
Co-reporter:Xiahong Xu, Zhou Nie, Jinhua Chen, Yingchun Fu, Wang Li, Qinpeng Shen and Shouzhuo Yao
Chemical Communications 2009(Issue 45) pp:NaN6948-6948
Publication Date(Web):2009/10/16
DOI:10.1039/B913943E
A novel label-free electrochemical strategy for monitoring the activity and inhibition of protein kinase is developed, based on the linkage between the phosphorylated peptide and DNA functionalized Au nanoparticles (DNA–AuNPs) by Zr4+ and the chronocoulometric response of [Ru(NH3)6]3+ absorbed on the DNA–AuNPs.
Co-reporter:Kaiyu He, Yong Li, Binbin Xiang, Peng Zhao, Yufang Hu, Yan Huang, Wang Li, Zhou Nie and Shouzhuo Yao
Chemical Science (2010-Present) 2015 - vol. 6(Issue 6) pp:NaN3564-3564
Publication Date(Web):2015/04/08
DOI:10.1039/C5SC00371G
Molecular logic gates are capable of performing various logic tasks for biomarker detection, disease diagnostics and therapy, and controlling biological progress. Herein, we integrated multiple components of a logic device into a single DNA 3D nano-assembly with a triangular prism structure. Compared with the separate construction of each component in previously reported DNA logic gate systems, such an integrated design strategy made the 3D DNA nanoprism universal for logic gates, it can be reconfigured to execute diverse logic operations. Binary basic logic gates (OR, AND, INHIBIT and XOR), combinatorial gates (INHIBIT–OR), and multi-valued logic gates (ternary INHIBIT gate) were readily achieved by taking this DNA nanoprism as a universal platform. Moreover, a logic gate system for identification of even numbers and odd numbers from natural numbers was established successfully by employing only this single DNA nanoprism and four short single-stranded DNA. The universality of this nanoprism greatly simplified the design of DNA logic gate system. Additionally, this nanoprism was able to perform logic operation steadily in a biological matrix, indicating that this box-like DNA nanostructure applies to logic gates in a complicated environment. This study provided a unique opportunity to design versatile 3D DNA nanostructure-based intelligent nanodevices, which show great potential in biocomputing, multi-parameter sensing, and intelligent disease diagnostics and therapy.
Co-reporter:Xiaohua Zhu, Mengjia Liu, Yang Liu, Ruwen Chen, Zhou Nie, Jinghong Li and Shouzhuo Yao
Journal of Materials Chemistry A 2016 - vol. 4(Issue 23) pp:NaN8977-8977
Publication Date(Web):2016/04/11
DOI:10.1039/C6TA01923D
Carbon coated hollow mesoporous FeP microcubes derived from Prussian blue were superior in catalytic activity and durability toward electrochemical hydrogen evolution with an overpotential of 115 mV to drive 10 mA cm−2.
Co-reporter:Shiyun Tang, Zhou Nie, Wang Li, Daiqi Li, Yan Huang and Shouzhuo Yao
Chemical Communications 2015 - vol. 51(Issue 76) pp:NaN14392-14392
Publication Date(Web):2015/08/03
DOI:10.1039/C5CC04170H
A label-free and convenient strategy for PARP-1 activity assay and inhibitors assessment has been developed based on the fluorescence resonance energy transfer (FRET) between a cationic conjugated polymer (CCP) and supercharged green fluorescent protein (scGFP).
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