研究者詳細 - 佐藤　しのぶ

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Specific metallization of double-stranded DNA using reducing group-labeled intercalator 査読有り国際誌

Himuro T., Araki R., Sato S., Takenaka S., Yasuda T.

IEEJ Transactions on Sensors and Micromachines ( 一般社団法人電気学会 ) 136 ( 10 ) 425 - 431 2016年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2016 The Institute of Electrical Engineers of Japan.We demonstrated that the DNA metallization technique using reducing group-labeled intercalator molecules permits dsDNA (double-stranded DNA) molecules to be specifically metallized while not permitting metallization of ssDNA (single-stranded DNA) molecules. First, dsDNA and ssDNA molecules were stretched on a mica surface using spin-coating technique, and metallization process was conducted. AFM observation showed that only dsDNA molecules were metallized, resulting in formation of nanowires with a diameter of about 11 nm. Secondly, dsDNA and ssDNA molecules were stretched and immobilized between two microelectrodes. Impedance analysis showed that the electrical conductivity of only dsDNA was drastically increased after the metallization process.

DOI： 10.1541/ieejsmas.136.425

CiNii Research

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2G17 部分的金属被覆DNAを用いたDNA分解酵素センサの開発査読有り

村上太理, 氷室貴大, 櫻田一歩, 佐藤しのぶ, 竹中繁織, 安田隆

バイオエンジニアリング講演会講演論文集 ( 一般社団法人日本機械学会 ) 2016.28 ( 0 ) _2G17-1_ - _2G17-3_ 2016年01月

記述言語：日本語掲載種別：研究論文（学術雑誌）

We developed a microdevice for detection of Deoxyribonuclease (DNase), which is a candidate biomarker for acute myocardial infarction. DNA metallization process using intercalator molecules labeled with reducing groups permits double-stranded DNA to be specifically metallized while not permitting metallization of single-stranded DNA. Using this process, we metallized DNA complexes consisting of single strands and double strands, and fabricated partially-metallized DNA nanowires. In this paper, multiple partially-metallized DNA nanowires were immobilized between two electrodes in a microchannel, and DNase detection was achieved by measuring the increase in impedance between the electrodes after DNA cleavage by DNase. The use of partially-metallized DNA nanowires having much smaller impedance compared to non-metallized DNA generates a drastic increase of impedance after their cleavage and therefore permits much higher sensitivity. The experimental results showed a definite correlation between DNase concentration and impedance increase rate in range of 10^<-6> to 10^<-1> unit/μl of DNase. The present study will make DNase detection more simple, rapid, sensitive than conventional fluorometric and electrochemical detections.

DOI： 10.1299/jsmebio.2016.28._2g17-1_

CiNii Research

その他リンク： https://www.jstage.jst.go.jp/article/jsmebio/2016.28/0/2016.28__2G17-1_/_pdf

A Selective G-Quadruplex DNA-Stabilizing Ligand Based on a Cyclic Naphthalene Diimide Derivative 査読有り国際誌

Md. Monirul Islam, Satoshi Fujii, Shinobu Sato, Tatsuo Okauchi & Shigeori Takenaka

Molecules 20 ( 6 ) 10963 - 10979 2015年06月

記述言語：英語掲載種別：研究論文（学術雑誌）

A cyclic naphthalene diimide (cyclic NDI, 1), carrying a benzene moiety as linker chain, was synthesized and its interaction with G-quadruplex DNAs of a-core and a-coreTT as a human telomeric DNA, c-kit and c-myc as DNA sequence at promoter region, or thrombin-binding aptamer (TBA) studied based on UV-VIS and circular dichroism (CD) spectroscopic techniques, thermal melting temperature measurement, and FRET-melting assay. The circular dichroism spectra showed that 1 induced the formation of different types of G-quadruplex DNA structure. Compound 1 bound to these G-quadruplexes with affinities in the range of 106 -107 M-1 order and a 2:1 stoichiometry. Compound 1 showed 270-fold higher selectivity for a-core than dsDNA with a preferable a-core binding than a-coreTT, c-kit, c-myc and TBA in the presence of K+ , which is supported by thermal melting studies. The FRET-melting assay also showed that 1 bound preferentially to human telomeric DNA. Compound 1 showed potent inhibition against telomerase activity with an IC<inf>50</inf> value of 0.9 μM and preferable binding to G-quadruplexes DNA than our previously published cyclic NDI derivative 3 carrying a benzene moiety as longer linker chain.

DOI： 10.3390/molecules200610963

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Erratum: Cooperative binding of ferrocenylnaphthalene diimide carrying β-cyclodextrin converts double-stranded DNA to a rod-like structure (Bioconjugate Chemistry (2015) 26:3 (379-382) DOI: 10.1021/bc500535n) 査読有り

Sato S., Umeda Y., Fujii S., Takenaka S.

Bioconjugate Chemistry 26 ( 4 ) 2015年04月

Cooperative Binding of Ferrocenylnaphthalene Diimide Carrying β-Cyclodextrin Converts Double-Stranded DNA to a Rod-Like Structure 査読有り国際誌

Shinobu Sato, Yuta Umeda, Satoshi Fujii, Shigeori Takenaka

Bioconjugate Chem. 26 ( 3 ) 379 - 382 2015年03月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

Ferrocenylnaphthalene diimide carrying β-cyclodextrin (β-CD), 1, intercalated into double-stranded DNA with a binding affinity of K = (6.6 ± 0.8) × 104 M-1 and a binding site size of n = 4, with a high positive cooperative parameter of ω = 14. β-CD and ferrocene moieties of the compound contributed to the formation of the intermolecular inclusion complex on DNA. Binding of 1 resulted in conversion of the DNA duplex to a rod-like form, which was cleaved upon adamantylamine addition. (Figure Presented).

DOI： 10.1021/bc500535n

その他リンク： http://pubs.acs.org/journal/bcches

Detection of deoxyribonuclease using site-specifically metallized DNA 査読有り

Himuro T., Murakami T., Sato S., Takenaka S., Yasuda T.

MicroTAS 2015 - 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences 1652 - 1654 2015年01月

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）

© 15CBMS-0001.We newly formed partially-metallized DNA nanowires, and used them for detection of deoxyribonuclease (DNase), enzyme for nonspecific DNA cleavage, which is a candidate biomarker for acute myocardial infarction. In this paper, we developed an original technique for site-specific metallization of DNA complexes consisting of single and double strands. Then, multiple DNA nanowires were immobilized between two electrodes in a microchannel, and DNase detection was achieved by measuring the increase in impedance between the electrodes after DNA cleavage by DNase. When DNase solutions of various concentrations were introduced, we succeeded in obtaining a definite correlation between DNase concentration and impedance increase rate in the range of 10-6 to 10-1 unit/μl.

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Ferrocenylnaphthalene Diimide-Based Electrochemical Detection of Aberrant Methylation in hTERT Gene 査読有り

Shinobu Sato, Toshiro Saeki, Tomoki Tanaka, Yusuke Kanezaki, Mana Hayakawa, Kazuya Haraguchi, Masaaki Kodama, Tatsuji Nishihara, Kazuhiro Tominaga, Shigeori Takenaka

Applied Biochemistry and Biotechnology 174 869 - 879 2014年07月

Metallization of double-stranded DNA triggered by bound galactose-modified naphthalene diimide 査読有り国際誌

Kohei Komizo , Hideyuki Ikedo , Shinobu Sato, Shigeori Takenaka

Bioconjugate Chemistry 25 ( 8 ) 1547 - 1555 2014年07月

記述言語：英語掲載種別：研究論文（学術雑誌）

Naphthalene diimide (NDI) derivatives bearing galactose moieties through different spacers, NDI-DS1 and NDI-DS2, were synthesized by the click reaction of the acetylene derivatives of NDI with galactose azide. They bound to double-stranded DNA with threading intercalation, as confirmed by the topoisomerase I assay and circular dichroism spectroscopy. The binding affinities of these ligands were on the order of 105 M-1 with several-fold higher affinity for double-stranded DNA than for single-stranded DNA. The silver mirror reaction on the double-stranded DNA bound to these ligands afforded silver nanowires that were converted to gold nanowires. In the atomic force microscopy measurements, the increased height of DNA areas on a mica plate was observed in the case of double-stranded DNA after NDI-DS2 treatment and subsequently silver mirror reaction, whereas the increased height of DNA areas was not observed in the case of single-stranded DNA after the same treatment. © 2014 American Chemical Society.

DOI： 10.1021/bc500263b

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Highly sensitive nuclease assay based on chemically-modified DNA or RNA 査読有り国際誌

Shinobu Sato, Shigeori Takenaka

Sensors 14 ( 7 ) 12437 - 12450 2014年07月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

Nucleolytic enzymes are associated with various diseases, and several methods have been developed for their detection. DNase expression is modulated in such diseases as acute myocardial infarction, transient myocardial ischemia, oral cancer, stomach cancer, and malignant lymphoma, and DNase I is used in cystic fibroma therapy. RNase is used to treat mesothelial cancer because of its antiproliferative, cytotoxic, and antineoplastic activities. Angiogenin, an angiogenic factor, is a member of the RNase A family. Angiogenin inhibitors are being developed as anticancer drugs. In this review, we describe fluorometric and electrochemical techniques for detecting DNase and RNase in disease. Oligonucleotides having fluorescence resonance energy transfer (FRET)-causing chromophores are non-fluorescent by themselves, yet become fluorescent upon cleavage by DNase or RNase. These oligonucleotides serve as a powerful tool to detect activities of these enzymes and provide a basis for drug discovery. In electrochemical techniques, ferrocenyl oligonucleotides with or without a ribonucleoside unit are used for the detection of RNase or DNase. This technique has been used to monitor blood or serum samples in several diseases associated with DNase and RNase and is unaffected by interferents in these sample types. © 2013 by the authors; licensee MDPI, Basel, Switzerland.

DOI： 10.3390/s140712437

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Design of tetraplex specific ligands: cyclic naphthalene diimide 査読有り国際誌

Yugo Esaki, Md. Monirul Islam, Satoshi Fujii, Shinobu Sato, Shigeori Takenaka

Chemical Communications 50 ( 45 ) 5967 - 5969 2014年04月

記述言語：英語掲載種別：研究論文（学術雑誌）

Cyclic naphthalene diimide 1 bound to hybrid-type tetraplex DNA from 5’-AGGG(TTAGGG)3-3’ (K = 8.6 × 106 M−1) with 260-fold greater affinity than binding to a double stranded oligonucleotide consisting of 5’-GGG AGG TTT CGC-3’ and 3’-CCC TCC AAA GCG-5’ (nK = 3.3 × 104 M−1) with 0.5 μM of IC50 for telomerase activity. © 2014 The Partner Organisations.

DOI： 10.1039/c4cc01005a

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Interactions of cyclic and non-cyclic naphthalene diimide derivativeswith different nucleic acids 査読有り国際誌

Izabella Czerwinska, Shinobu Sato, Bernard Juskowiak, Shigeori Takenaka

Bioorganic & Medicinal Chemistry 22 ( 9 ) 2593 - 2601 2014年03月

記述言語：英語掲載種別：研究論文（学術雑誌）

Recently, strategy based on stabilization of G-quadruplex telomeric DNA by small organic molecule has been realized by naphthalene diimide derivatives (NDIs). At the same time NDIs bind to DNA duplex as threading intercalators. Here we present cyclic derivative of naphthalene diimide (ligand 1) as DNA-binding ligand with ability to recognition of different structures of telomeric G-quadruplexes and ability to bis-intercalate to double-stranded helixes. The results have been compared to non-cyclic derivative (ligand 2) and revealed that preferential binding of ligands to nucleic acids strongly depends on their topology and structural features of ligands. © 2014 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.bmc.2014.03.034

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ヒトテロメアDNAを用いたナトリウムイオン蛍光イメージング試薬（SSO）の開発

今市悠貴, 古賀晴香, 中澤浩二, 佐藤しのぶ, 竹中繁織

ナノ学会会報 ( ナノ学会 ) 12 73 - 77 2014年02月

ヒトテロメアDNAを用いたナトリウムイオン蛍光イメージング試薬（SSO）の開発" 今市悠貴, 古賀晴香, 中澤浩二, 佐藤しのぶ, 竹中繁織招待有り査読有り

今市悠貴, 古賀晴香, 中澤浩二, 佐藤しのぶ, 竹中繁織

ナノ学会会報 12 ( 2 ) 73 - 77 2014年02月

Electrochemical telomerase assay for oral cancer screening 査読有り国際誌

Tominaga K., Hayakawa M., Sato S., Kodama M., Habu M., Takenaka S.

World Automation Congress Proceedings 38 - 42 2014年01月

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）

© 2014 TSI Press.Telomerase has long been known as a cancer marker. As an easy-to-perform oral cancer screening system, we established a new electrochemical telomerase assay (ECTA) method that is superior to a telomerase repeat amplification protocol assay, a popular method for detection of telomerase activity. In the present study, we employed 3 types of clinical samples to examine the sensitivity and specificity of ECTA. Three types of samples were obtained from 44 oral squamous cell carcinoma patients and 16 healthy volunteers; exfoliated cells from the whole oral cavity, exfoliated cells from local lesions, and tissue samples from lesions. Telomerase activity was determined in the samples using ECTA. The increase in current in the oral cancer group was significantly higher than that in the healthy volunteer group for each sample type, while there were no significant differences among the 3 sample types in the oral cancer group. The sensitivity and specificity of ECTA was 94.6% and 88.6%, respectively. Our novel ECTA method showed excellent detection of telomerase activity and is consider applicable as an oral cancer screening system.

DOI： 10.1109/WAC.2014.6935647

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1C14 2本鎖DNAの特異的な金属被覆によるナノワイヤの形成とその電気的特性の評価(OS14-1:バイオMEMS(1))

荒木遼, 氷室貴大, 池堂英幸, 佐藤しのぶ, 竹中繁織, 安田隆

バイオエンジニアリング講演会講演論文集 ( 一般社団法人日本機械学会 ) 2014 ( 0 ) 73 - 74 2014年01月

Development of a membrane-based microwave mediated electrochemical ELISA method for TNF-α detection in patients with periodontitis 査読有り国際誌

Irmina DIALA, Shinobu SATO, Michihiko USUI, Keisuke NAKASHIMA, Tatsuji NISHIHARA, Shigeori TAKENAKA

Analytical Sciences 29 ( 9 ) 927 - 930 2013年09月

記述言語：英語掲載種別：研究論文（学術雑誌）

The tumor necrosis factor-α (TNF-α) is implicated in periodontal disease, and there was an attempt to quantitate it by a membrane-based microwave-mediated electrochemical enzyme-linked immunosorbent assay (MMeELISA) using p-aminophenyl phosphate (pAPP) with an over-all time of 1.5 h. A differential pulse voltammetric signal increased linearly with an increase in the amount of TNF-α with a detection limit of 0.48 pg mL-1. This bio-sensing platform revealed a significant difference in the TNF-α level between GCF samples from periodontal patients and healthy volunteers. © 2013 The Japan Society for Analytical Chemistry.

DOI： 10.2116/analsci.29.927

その他リンク： https://www.jstage.jst.go.jp/article/analsci/29/9/29_927/_pdf

Electrochemical Detection of Duplex DNA Using Intercalation-Triggered Decomplexation of Ferrocene with β-Cyclodextrin 査読有り国際誌

Hirotomo Takenaka, Shinobu Sato, Shigeori Takenaka

Electroanalysis 25 ( 8 ) 1827 - 1830 2013年08月

記述言語：英語掲載種別：研究論文（学術雑誌）

The redox peak of ferrocenylnaphthalene diimide used as a threading intercalator shifted positively due to the formation of its complex with β-cyclodextrin. This complex collapsed upon the addition of double-stranded DNA, and its redox potential shifted negatively. This behavior was applied for the homogenous detection of a polymerase chain reaction (PCR) product from Porphyromonas gingivalis, which is important for the diagnosis of periodontal disease, and its quantitative detection was achieved with a detection limit of 2.7nM. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/elan.201300211

その他リンク： http://onlinelibrary.wiley.com/doi/10.1002/elan.201300211/abstract

Naphthalene diimide carrying two cysteine termini at both imide linkers as a molecular staple 査読有り国際誌

Shinobu Sato, Kosuke Yamamura, Shigeori Takenaka

Electroanalysis 25 ( 8 ) 1831 - 1839 2013年08月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

Naphthalene diimide (1) carrying cysteines at the termini of amide substituents were synthesized to act as a molecular staple of double stranded DNA. Since 1 is able to bind to double stranded DNA with threading intercalation, the complex of 1 with double stranded DNA can be topologically immobilized on a gold surface through the S-Au linkage as confirmed by cyclic voltammetric experiment. Ferrocenyl-double stranded 23-mertic oligonucleotide, dsFcODN, was immobilized on gold electrode with 1.0×1012 molecules cm-2 when electrode was treated with 2.0μM dsFcODN and 4.0μM 1 for 1h at room temperature. The coverage density was similar to that obtained for the terminal thiol-modified oligonucleotide. Compound 1 was applied to detect the 321-meric PCR product of P. gingivalis, which is important in the diagnosis of periodontal disease. This experiment, coupled with the use of ferrocenylnaphthalene diimide, FND as electrochemical indicator for double stranded DNA, resulted in quantitative detection of PCR product within the range of 10pgμL-1-10ngμL-1 (15nM-15μM). The 1 and FND established a simple and rapid detection method of double stranded PCR product with a detection limit of 10pgμL-1 (15nM). © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DOI： 10.1002/elan.201300209

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電気化学的遺伝子検出における超分子形成査読有り

Shinobu Sato, Shigeori Takenaka

BUNSEKI KAGAKU 62 ( 7 ) 627 - 635 2013年07月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

Gene detection based on electrochemical techniques has been gaining much attention in recent years, not only because it provides simple and rapid methods for measurement and detec t ion, but al so for the eas ines s of downs i z ing the ins t rumentat ion. Ferrocenylnaphthalene diimide, which forms a pseudo-catenane complex with double-stranded DNA, has been developed to be used in an electrochemical detection system for doublestranded DNA coupled with a DNA probe-immobilized electrode. To improve the performance of electrochemical gene detection, FND or its derivatives, which can form a supramolecular complex, have been under development. Here, we review an electrochemical DNA or DNA-related enzyme detection technique based on supramolecular complex formation. © 2013 The Japan Society for Analytical Chemistry.

DOI： 10.2116/bunsekikagaku.62.627