Updated on 2024/09/23

写真a

 
HAYASHI Gosuke
 
Organization
Graduate School of Engineering Biomolecular Engineering 1 Associate professor
Graduate School
Graduate School of Engineering
Undergraduate School
School of Engineering Chemistry and Biotechnology
Title
Associate professor
Contact information
メールアドレス
External link

Degree 1

  1. 博士(理学) ( 2009.3   大阪大学 ) 

Research Interests 3

  1. Peptide chemistry

  2. Chemical protein synthesis

  3. Epigenetics

Research Areas 3

  1. Nanotechnology/Materials / Bio chemistry

  2. Nanotechnology/Materials / Chemistry and chemical methodology of biomolecules

  3. Nanotechnology/Materials / Bio chemistry

Research History 13

  1. Nagoya University   Graduate School of Engineering, Department of Bimolecular Engineering   Associate professor

    2019.3

      More details

    Country:Japan

  2. Nagoya University   Graduate School of Engineering, Department of Bimolecular Engineering   Associate professor

    2019.3

  3. Nagoya University   Graduate School of Engineering Biomolecular Engineering 1   Associate professor

    2019.3

  4. The University of Tokyo   Graduate School of Engineering, Department of Chemistry and Biotechnology   Associate Professor

    2013.2 - 2019.2

      More details

    Country:Japan

  5. The University of Tokyo   Graduate School of Engineering, Department of Chemistry and Biotechnology   Assistant Professor

    2013.2 - 2019.2

  6. The University of Tokyo   Research Center for Advanced Science and Technology   Associate Professor

    2012.6 - 2013.1

      More details

    Country:Japan

  7. The University of Tokyo   Research Center for Advanced Science and Technology   Designated assistant professor

    2012.6 - 2013.1

  8. Boston University   Department of Biomedical Engineering   Postdoctral Fellow (JSPS Reserch Fellowship for Young Scientists PD and Overseas Research Fellowships)

    2011.2 - 2012.5

      More details

    Country:Japan

  9. Boston University   Department of Biomedical Engineering   Postdoctral Fellow (JSPS Reserch Fellowship for Young Scientists PD and Overseas Research Fellowships)

    2011.2 - 2012.5

  10. The University of Tokyo   Research Center for Advanced Science and Technology   Postdoctral Fellow (JSPS Research Fellowship for Young Scientists PD)

    2009.4 - 2011.2

      More details

    Country:Japan

  11. The University of Tokyo   Research Center for Advanced Science and Technology   Postdoctral Fellow (JSPS Research Fellowship for Young Scientists PD)

    2009.4 - 2011.2

  12. Osaka University   Graduate School of Science, Department of Chemistry   JSPS Research Fellowship of Young Scientists DC1

    2006.4 - 2009.3

      More details

    Country:Japan

  13. Osaka University   Graduate School of Science, Department of Chemistry   JSPS Research Fellowship of Young Scientists DC1

    2006.4 - 2009.3

▼display all

Education 6

  1. Osaka University   Graduate School, Division of Natural Science   Department of Chemistry

    2006.4 - 2009.3

      More details

    Country: Japan

  2. Osaka University   Graduate School, Division of Natural Science   Department of Chemistry

    2006.4 - 2009.3

      More details

    Country: Japan

  3. Kyoto University   Graduate School, Division of Engineering   Department of Synthetic Chemistry and Biological Chemistry

    2004.4 - 2006.3

      More details

    Country: Japan

  4. Kyoto University   Graduate School, Division of Engineering   Department of Synthetic Chemistry and Biological Chemistry

    2004.4 - 2006.3

      More details

    Country: Japan

  5. Kyoto University   Faculty of Engineering   Undergraduate School of Industrial Chemistry

    2000.4 - 2004.3

      More details

    Country: Japan

  6. Kyoto University   Faculty of Engineering   Undergraduate School of Industrial Chemistry

    2000.4 - 2004.3

      More details

    Country: Japan

▼display all

Professional Memberships 10

  1. 日本化学会

  2. 日本生化学会

  3. 日本分子生物学会

  4. 日本ケミカルバイオロジー学会

  5. 日本エピジェネティクス研究会

  6. 日本生化学会

  7. 日本化学会

  8. 日本分子生物学会

  9. 日本ケミカルバイオロジー学会

  10. 日本エピジェネティクス研究会

▼display all

Awards 16

  1. 有機合成化学奨励賞

    2021   有機合成化学協会  

  2. 若い世代の特別講演賞

    2020.3   日本化学会  

     More details

    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  3. 若い世代の特別講演賞

    2020.3   日本化学会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  4. 有機合成化学協会 研究企画賞

    2020   有機合成化学協会  

  5. 日本ペプチド学会 奨励賞

    2019.11   日本ペプチド学会  

     More details

    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  6. 日本ペプチド学会 奨励賞

    2019.11   日本ペプチド学会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  7. 日本エピジェネティクス研究会 奨励賞

    2018.3   日本エピジェネティクス研究会  

     More details

    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  8. 日本エピジェネティクス研究会 奨励賞

    2018.3   日本エピジェネティクス研究会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  9. バイオ関連化学シンポジウム 講演賞

    2017.9   日本化学会  生体機能関連化学部会、バイオテクノロジー部会  

     More details

    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  10. バイオ関連化学シンポジウム 講演賞

    2017.9   日本化学会 生体機能関連化学部会、バイオテクノロジー部会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  11. 日本化学会優秀講演賞(学術)

    2017.3   日本化学会  

     More details

    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  12. 日本化学会優秀講演賞(学術)

    2017.3   日本化学会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  13. Outstanding Young Researcher Poster Presentation Award

    2010.12   PACIFICHEM2010  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  14. Outstanding Young Researcher Poster Presentation Award

    2010.12   PACIFICHEM2010  

     More details

    Award type:International academic award (Japan or overseas)  Country:Japan

  15. Poster Award in 4th International Symposium on Nucleic Acids Chemistry

    2005.9   日本核酸化学会  

     More details

    Award type:Award from international society, conference, symposium, etc.  Country:Japan

  16. Poster Award in 4th International Symposium on Nucleic Acids Chemistry

    2005.9   日本核酸化学会  

     More details

    Award type:International academic award (Japan or overseas)  Country:Japan

▼display all

 

Papers 66

  1. H3K4me1 recruits DNA repair proteins in plants

    Quiroz, D; Oya, S; Lopez-Mateos, D; Zhao, K; Pierce, A; Ortega, L; Ali, A; Carbonell-Bejerano, P; Yarov-Yarovoy, V; Suzuki, S; Hayashi, G; Osakabe, A; Monroe, G

    PLANT CELL   Vol. 36 ( 6 ) page: 2410 - 2426   2024.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Plant Cell  

    DNA repair proteins can be recruited by their histone reader domains to specific epigenomic features, with consequences on intragenomic mutation rate variation. Here, we investigated H3K4me1-associated hypomutation in plants. We first examined 2 proteins which, in plants, contain Tudor histone reader domains: PRECOCIOUS DISSOCIATION OF SISTERS 5 (PDS5C), involved in homology-directed repair, and MUTS HOMOLOG 6 (MSH6), a mismatch repair protein. The MSH6 Tudor domain of Arabidopsis (Arabidopsis thaliana) binds to H3K4me1 as previously demonstrated for PDS5C, which localizes to H3K4me1-rich gene bodies and essential genes. Mutations revealed by ultradeep sequencing of wild-type and msh6 knockout lines in Arabidopsis show that functional MSH6 is critical for the reduced rate of single-base substitution (SBS) mutations in gene bodies and H3K4me1-rich regions. We explored the breadth of these mechanisms among plants by examining a large rice (Oryza sativa) mutation data set. H3K4me1-associated hypomutation is conserved in rice as are the H3K4me1-binding residues of MSH6 and PDS5C Tudor domains. Recruitment of DNA repair proteins by H3K4me1 in plants reveals convergent, but distinct, epigenome-recruited DNA repair mechanisms from those well described in humans. The emergent model of H3K4me1-recruited repair in plants is consistent with evolutionary theory regarding mutation modifier systems and offers mechanistic insight into intragenomic mutation rate variation in plants.

    DOI: 10.1093/plcell/koae089

    Web of Science

    Scopus

    PubMed

  2. Optineurin provides a mitophagy contact site for TBK1 activation

    Yamano, K; Sawada, M; Kikuchi, R; Nagataki, K; Kojima, W; Endo, R; Kinefuchi, H; Sugihara, A; Fujino, T; Watanabe, A; Tanaka, K; Hayashi, G; Murakami, H; Matsuda, N

    EMBO JOURNAL   Vol. 43 ( 5 ) page: 754 - 779   2024.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:EMBO Journal  

    Tank-binding kinase 1 (TBK1) is a Ser/Thr kinase that is involved in many intracellular processes, such as innate immunity, cell cycle, and apoptosis. TBK1 is also important for phosphorylating the autophagy adaptors that mediate the selective autophagic removal of damaged mitochondria. However, the mechanism by which PINK1-Parkin-mediated mitophagy activates TBK1 remains largely unknown. Here, we show that the autophagy adaptor optineurin (OPTN) provides a unique platform for TBK1 activation. Both the OPTN-ubiquitin and the OPTN-pre-autophagosomal structure (PAS) interaction axes facilitate assembly of the OPTN-TBK1 complex at a contact sites between damaged mitochondria and the autophagosome formation sites. At this assembly point, a positive feedback loop for TBK1 activation is initiated that accelerates hetero-autophosphorylation of the protein. Expression of monobodies engineered here to bind OPTN impaired OPTN accumulation at contact sites, as well as the subsequent activation of TBK1, thereby inhibiting mitochondrial degradation. Taken together, these data show that a positive and reciprocal relationship between OPTN and TBK1 initiates autophagosome biogenesis on damaged mitochondria.

    DOI: 10.1038/s44318-024-00036-1

    Web of Science

    Scopus

    PubMed

  3. One-pot ligation of multiple peptide segments via N-terminal thiazolidine deprotection chemistry.

    Nakamura G, Nakatsu K, Hayashi G

    Methods in enzymology   Vol. 698   page: 169 - 194   2024

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Methods in Enzymology  

    Peptide ligation chemistries have revolutionized the synthesis of proteins with site-specific modifications or proteomimetics through assembly of multiple peptide segments. In order to prepare polypeptide chains consisting of 100–150 amino acid residues or larger generally assembled from three or more peptide segments, iterative purification process that decreases the product yield is usually demanded. Accordingly, methodologies for one-pot peptide ligation that omit the purification steps of intermediate peptide segments have been vigorously developed so far to improve the efficiency of chemical protein synthesis. In this chapter, we first outline the concept and recent advances of one-pot peptide ligation strategies. Then, the practical guideline for the preparation of peptide segments for one-pot peptide ligation is described with an emphasis on diketopiperazine thioester synthesis. Finally, we disclose the explicit protocols for one-pot four segment ligation via repetitive deprotection of N-terminal thiazolidine by a 2-aminobenzamide type aldehyde scavenger.

    DOI: 10.1016/bs.mie.2024.04.020

    Scopus

    PubMed

  4. Contributions of histone tail clipping and acetylation in nucleosome transcription by RNA polymerase II

    Oishi, T; Hatazawa, S; Kujirai, T; Kato, J; Kobayashi, Y; Ogasawara, M; Akatsu, M; Ehara, H; Sekine, SI; Hayashi, G; Takizawa, Y; Kurumizaka, H

    NUCLEIC ACIDS RESEARCH   Vol. 51 ( 19 ) page: 10364 - 10374   2023.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nucleic Acids Research  

    The N-terminal tails of histones protrude from the nucleosome core and are target sites for histone modifications, such as acetylation and methylation. Histone acetylation is considered to enhance transcription in chromatin. However, the contribution of the histone N-terminal tail to the nucleosome transcription by RNA polymerase II (RNAPII) has not been clarified. In the present study, we reconstituted nucleosomes lacking the N-terminal tail of each histone, H2A, H2B, H3 or H4, and performed RNAPII transcription assays. We found that the N-terminal tail of H3, but not H2A, H2B and H4, functions in RNAPII pausing at the SHL(-5) position of the nucleosome. Consistently, the RNAPII transcription assay also revealed that the nucleosome containing N-terminally acetylated H3 drastically alleviates RNAPII pausing at the SHL(-5) position. In addition, the H3 acetylated nucleosome produced increased amounts of the run-off transcript. These results provide important evidence that the H3 N-terminal tail plays a role in RNAPII pausing at the SHL(-5) position of the nucleosome, and its acetylation directly alleviates this nucleosome barrier.

    DOI: 10.1093/nar/gkad754

    Web of Science

    Scopus

    PubMed

  5. Large-scale analysis of mRNA sequences localized near the start and amber codons and their impact on the diversity of mRNA display libraries

    Umemoto, S; Kondo, T; Fujino, T; Hayashi, G; Murakami, H

    NUCLEIC ACIDS RESEARCH   Vol. 51 ( 14 ) page: 7465 - 7479   2023.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nucleic Acids Research  

    Extremely diverse libraries are essential for effectively selecting functional peptides or proteins, and mRNA display technology is a powerful tool for generating such libraries with over 1012-1013 diversity. Particularly, the protein-puromycin linker (PuL)/mRNA complex formation yield is determining for preparing the libraries. However, how mRNA sequences affect the complex formation yield remains unclear. To study the effects of N-terminal and C-terminal coding sequences on the complex formation yield, puromycin-attached mRNAs containing three random codons after the start codon (32768 sequences) or seven random bases next to the amber codon (6480 sequences) were translated. Enrichment scores were calculated by dividing the appearance rate of every sequence in protein-PuL/mRNA complexes by that in total mRNAs. The wide range of enrichment scores (0.09-2.10 for N-terminal and 0.30-4.23 for C-terminal coding sequences) indicated that the N-terminal and C-terminal coding sequences strongly affected the complex formation yield. Using C-terminal GGC-CGA-UAG-U sequences, which resulted in the highest enrichment scores, we constructed highly diverse libraries of monobodies and macrocyclic peptides. The present study provides insights into how mRNA sequences affect the protein/mRNA complex formation yield and will accelerate the identification of functional peptides and proteins involved in various biological processes and having therapeutic applications.

    DOI: 10.1093/nar/gkad555

    Web of Science

    Scopus

    PubMed

  6. Thiocholine-Mediated One-Pot Peptide Ligation and Desulfurization

    Suzuki, S; Nakajima, Y; Kamo, N; Osakabe, A; Okamoto, A; Hayashi, G; Murakami, H

    MOLECULES   Vol. 28 ( 9 )   2023.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Molecules  

    Thiol catalysts are essential in native chemical ligation (NCL) to increase the reaction efficiency. In this paper, we report the use of thiocholine in chemical protein synthesis, including NCL-based peptide ligation and metal-free desulfurization. Evaluation of thiocholine peptide thioester in terms of NCL and hydrolysis kinetics revealed its practical utility, which was comparable to that of other alkyl thioesters. Importantly, thiocholine showed better reactivity as a thiol additive in desulfurization, which is often used in chemical protein synthesis to convert Cys residues to more abundant Ala residues. Finally, we achieved chemical synthesis of two differently methylated histone H3 proteins via one-pot NCL and desulfurization with thiocholine.

    DOI: 10.3390/molecules28093655

    Web of Science

    Scopus

    PubMed

  7. Structural basis for the unique multifaceted interaction of DPPA3 with the UHRF1 PHD finger.

    Hata K, Kobayashi N, Sugimura K, Qin W, Haxholli D, Chiba Y, Yoshimi S, Hayashi G, Onoda H, Ikegami T, Mulholland CB, Nishiyama A, Nakanishi M, Leonhardt H, Konuma T, Arita K

    Nucleic acids research   Vol. 50 ( 21 ) page: 12527 - 12542   2022.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nar/gkac1082

    PubMed

  8. Structural basis for the unique multifaceted interaction of DPPA3 with the UHRF1 PHD finger

    Hata K., Kobayashi N., Sugimura K., Qin W., Haxholli D., Chiba Y., Yoshimi S., Hayashi G., Onoda H., Ikegami T., Mulholland C.B., Nishiyama A., Nakanishi M., Leonhardt H., Konuma T., Arita K.

    Nucleic acids research   Vol. 50 ( 21 ) page: 12527 - 12542   2022.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nucleic acids research  

    Ubiquitin-like with PHD and RING finger domain-containing protein 1 (UHRF1)-dependent DNA methylation is essential for maintaining cell fate during cell proliferation. Developmental pluripotency-associated 3 (DPPA3) is an intrinsically disordered protein that specifically interacts with UHRF1 and promotes passive DNA demethylation by inhibiting UHRF1 chromatin localization. However, the molecular basis of how DPPA3 interacts with and inhibits UHRF1 remains unclear. We aimed to determine the structure of the mouse UHRF1 plant homeodomain (PHD) complexed with DPPA3 using nuclear magnetic resonance. Induced α-helices in DPPA3 upon binding of UHRF1 PHD contribute to stable complex formation with multifaceted interactions, unlike canonical ligand proteins of the PHD domain. Mutations in the binding interface and unfolding of the DPPA3 helical structure inhibited binding to UHRF1 and its chromatin localization. Our results provide structural insights into the mechanism and specificity underlying the inhibition of UHRF1 by DPPA3.

    DOI: 10.1093/nar/gkac1082

    Scopus

  9. Structural basis for the unique multifaceted interaction of DPPA3 with the UHRF1 PHD finger

    Hata Keiichi, Kobayashi Naohiro, Sugimura Keita, Qin Weihua, Haxholli Deis, Chiba Yoshie, Yoshimi Sae, Hayashi Gosuke, Onoda Hiroki, Ikegami Takahisa, Mulholland Christopher B., Nishiyama Atsuya, Nakanishi Makoto, Leonhardt Heinrich, Konuma Tsuyoshi, Arita Kyohei

    NUCLEIC ACIDS RESEARCH   Vol. 50 ( 21 ) page: 12527 - 12542   2022.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nar/gkac1082

    Web of Science

  10. Structural basis for the unique multifaceted interaction of DPPA3 with the UHRF1 PHD finger

    Hata Keiichi, Kobayashi Naohiro, Sugimura Keita, Qin Weihua, Haxholli Deis, Chiba Yoshie, Yoshimi Sae, Hayashi Gosuke, Onoda Hiroki, Ikegami Takahisa, Mulholland Christopher B., Nishiyama Atsuya, Nakanishi Makoto, Leonhardt Heinrich, Konuma Tsuyoshi, Arita Kyohei

    NUCLEIC ACIDS RESEARCH     2022.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nar/gkac1082

    Web of Science

  11. Repetitive Thiazolidine Deprotection Using a Thioester-Compatible Aldehyde Scavenger for One-Pot Multiple Peptide Ligation. International journal

    Koki Nakatsu, Akimitsu Okamoto, Gosuke Hayashi, Hiroshi Murakami

    Angewandte Chemie (International ed. in English)   Vol. 61 ( 39 ) page: e202206240   2022.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Strategies for one-pot peptide ligation enable chemists to access synthetic proteins at a high yield in a short time. Herein, we report a novel one-pot multi-segments ligation strategy using N-terminal thiazolidine (Thz) peptide and a newly designed formaldehyde scavenger. Among the designed 2-aminobenzamide-based aldehyde scavengers, 2-amino-5-methoxy-N',N'-dimethylbenzohydrazide (AMDBH) can remarkably convert Thz into unprotected cysteine at pH 4.0. Furthermore, AMDBH degrades Thz at a considerably low rate at pH 7.5, and thioester degradation caused by this scavenger is negligible. As a result, we have developed an efficient one-pot peptide ligation strategy by simply repetitively changing the pH with AMDBH. Finally, we synthesize mono-ubiquitinated histone H2A.Z (209 amino acids) via AMDBH-mediated one-pot four-segment peptide ligation in good yield.

    DOI: 10.1002/anie.202206240

    Web of Science

    Scopus

    PubMed

  12. One-pot peptide ligation mediated by 2-aminobenzamide-type aldehyde scavengers

    Nakatsu, K; Okamoto, A; Murakami, H; Hayashi, G

    JOURNAL OF PEPTIDE SCIENCE   Vol. 28   2022.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Web of Science

  13. Peptide selenoester synthesis via diketopiperazine formation

    Hayashi, G; Hashimoto, M; Nakatsu, K; Murakami, H

    JOURNAL OF PEPTIDE SCIENCE   Vol. 28   2022.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Web of Science

  14. Monobodies with potent neutralizing activity against SARS-CoV-2 Delta and other variants of concern

    Kondo, T; Matsuoka, K; Umemoto, S; Fujino, T; Hayashi, G; Iwatani, Y; Murakami, H

    LIFE SCIENCE ALLIANCE   Vol. 5 ( 6 )   2022.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Life Science Alliance  

    Neutralizing antibodies against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are useful for patients’ treatment of the coronavirus disease 2019 (COVID-19). We report here affinity maturation of monobodies against the SARS-CoV-2 spike protein and their neutralizing activity against SARS-CoV-2 B.1.1 (Pango v.3.1.14) as well as four variants of concern. We selected matured monobodies from libraries with multi-site saturation mutagenesis on the recognition loops through in vitro selection. One clone, the C4-AM2 monobody, showed extremely high affinity (KD < 0.01 nM) against the receptor-binding domain of the SARS-CoV-2 B.1.1, even in monomer form. Furthermore, the C4-AM2 monobody efficiently neutralized the SARS-CoV-2 B.1.1 (IC50 = 46 pM, 0.62 ng/ml), and the Alpha (IC50 = 77 pM, 1.0 ng/ml), Beta (IC50 = 0.54 nM, 7.2 ng/ml), Gamma (IC50 = 0.55 nM, 7.4 ng/ml), and Delta (IC50 = 0.59 nM, 8.0 ng/ml) variants. The obtained monobodies would be useful as neutralizing proteins against current and potentially hazardous future SARS-CoV-2 variants.

    DOI: 10.26508/lsa.202101322

    Web of Science

    Scopus

    PubMed

    CiNii Research

  15. Construction of a Highly Diverse mRNA Library for <i>in vitro</i> Selection of Monobodies

    Kondo, T; Eguchi, M; Tsuzuki, N; Murata, N; Fujino, T; Hayashi, G; Murakami, H

    BIO-PROTOCOL   Vol. 11 ( 16 ) page: e4125   2021.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Bio-protocol  

    Recently, we developed transcription/translation coupled with the association of puromycin linker (TRAP) display as a quick in vitro selection method to obtain antibody-like proteins. For the in vitro selection, it is important to prepare mRNA libraries among which the diversity is high. Here, we describe a method for the preparation of monobody mRNA libraries with greater than 1013 theoretical diversity. First, we synthesized two long single-stranded DNAs that corresponded to fragments of monobody DNA, with random codons in the BC and FG loops. These oligonucleotides were ligated by T4 DNA ligase with the support of guide oligonucleotides containing 3′ ends that were protected by a modification. After amplifying the product DNAs by PCR, one end of each DNA fragment was digested with the type II restriction enzyme BsaI, and the resulting DNA fragments were ligated using T4 DNA ligase. After amplification of the DNA product, mRNAs were synthesized by T7 RNA polymerase. This method is simple and could be used for the preparation of mRNA libraries for various antibody-like proteins.

    DOI: 10.21769/BioProtoc.4125

    Web of Science

    Scopus

    PubMed

  16. Base-resolution analysis of 5-hydroxymethylcytidine by selective oxidation and reverse transcription arrest

    Koyama, K; Hayashi, G; Ueda, H; Ota, S; Nagae, G; Aburatani, H; Okamoto, A

    ORGANIC & BIOMOLECULAR CHEMISTRY   Vol. 19 ( 29 ) page: 6478 - 6486   2021.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Organic and Biomolecular Chemistry  

    While 5-hydroxymethylcytidine in RNA (hm5C) is associated with cellular development and differentiation, its distribution and biological function remain largely unexplored because suitable detection methods are lacking. Here, we report a base-resolution sequencing method forhm5C in RNA by applying peroxotungstate-mediated chemical conversion ofhm5C to trihydroxylated thymine (thT). Reverse transcription by SuperScript III terminated at thethT site, probably because of its unnatural nucleobase structure producing truncated cDNA. Consequently, base-resolution analysis of thehm5C sites in RNA was achieved with both Sanger sequencing and Illumina sequencing analysis by comparing sequencing data before and after peroxotungstate treatment.

    DOI: 10.1039/d1ob00995h

    Web of Science

    Scopus

    PubMed

  17. Silyl-protected propargyl glycine for multiple labeling of peptides by chemoselective silyl-deprotection

    Naoki Kamo, Gosuke Hayashi, Akimitsu Okamoto

    Tetrahedron Letters   Vol. 73   2021.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier Ltd  

    We synthesized Fmoc-propargyl glycine derivatives bearing different silyl protecting groups that can be readily introduced by using a standard solid-phase peptide coupling procedures. Taking advantage of the orthogonality between the different silyl protecting groups, chemoselective incorporation of functional molecules into a 19-mer peptide through click reactions was demonstrated.

    DOI: 10.1016/j.tetlet.2021.153093

    Web of Science

    Scopus

  18. Organoruthenium-catalyzed chemical protein synthesis to elucidate the functions of epigenetic modifications on heterochromatin factors†

    Kamo, N; Kujirai, T; Kurumizaka, H; Murakami, H; Hayashi, G; Okamoto, A

    CHEMICAL SCIENCE   Vol. 12 ( 16 ) page: 5926 - 5937   2021.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Chemical Science  

    The application of organometallic compounds for protein science has received attention. Recently, total chemical protein synthesis using transition metal complexes has been developed to produce various proteins bearing site-specific posttranslational modifications (PTMs). However, in general, significant amounts of metal complexes were required to achieve chemical reactions of proteins bearing a large number of nucleophilic functional groups. Moreover, syntheses of medium-size proteins (>20 kDa) were plagued by time-consuming procedures due to cumbersome purification and isolation steps, which prevented access to variously decorated proteins. Here, we report a one-pot multiple peptide ligation strategy assisted by an air-tolerant organoruthenium catalyst that showed more than 50-fold activity over previous palladium complexes, leading to rapid and quantitative deprotection on a protein with a catalytic amount (20 mol%) of the metal complex even in the presence of excess thiol moieties. Utilizing the organoruthenium catalyst, heterochromatin factors above 20 kDa, such as linker histone H1.2 and heterochromatin protein 1α (HP1α), bearing site-specific PTMs including phosphorylation, ubiquitination, citrullination, and acetylation have been synthesized. The biochemical assays using synthetic proteins revealed that the citrullination at R53 in H1.2 resulted in the reduced electrostatic interaction with DNA and the reduced binding affinity to nucleosomes. Furthermore, we identified a key phosphorylation region in HP1α to control its DNA-binding ability. The ruthenium chemistry developed here will facilitate the preparation of a variety of biologically and medically significant proteins containing PTMs and non-natural amino acids.

    DOI: 10.1039/d1sc00731a

    Web of Science

    Scopus

    PubMed

  19. cDNA TRAP display for rapid and stable <i>in vitro</i> selection of antibody-like proteins

    Kondo, T; Eguchi, M; Kito, S; Fujino, T; Hayashi, G; Murakami, H

    CHEMICAL COMMUNICATIONS   Vol. 57 ( 19 ) page: 2416 - 2419   2021.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Chemical Communications  

    We developed a cDNA TRAP display for the rapid selection of antibody-like proteins in various conditions. By modifying the original puromycin linker in the TRAP display, a monobody was covalently attached to the cDNA. As a proof-of-concept, we demonstrated a rapid model selection of an anti-EGFR1 monobody in a solution containing ribonuclease.

    DOI: 10.1039/d0cc07541h

    Web of Science

    Scopus

    PubMed

  20. Acetylation-modulated communication between the H3 N-terminal tail domain and the intrinsically disordered H1 C-terminal domain

    Hao, FF; Murphy, KJ; Kujirai, T; Kamo, N; Kato, J; Koyama, M; Okamato, A; Hayashi, G; Kurumizaka, H; Hayes, JJ

    NUCLEIC ACIDS RESEARCH   Vol. 48 ( 20 ) page: 11510 - 11520   2020.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nucleic Acids Research  

    Linker histones (H1s) are key structural components of the chromatin of higher eukaryotes. However, the mechanisms by which the intrinsically disordered linker histone carboxy-terminal domain (H1 CTD) influences chromatin structure and gene regulation remain unclear. We previously demonstrated that the CTD of H1.0 undergoes a significant condensation (reduction of end-to-end distance) upon binding to nucleosomes, consistent with a transition to an ordered structure or ensemble of structures. Here, we show that deletion of the H3 N-terminal tail or the installation of acetylation mimics or bona fide acetylation within H3 N-terminal tail alters the condensation of the nucleosome-bound H1 CTD. Additionally, we present evidence that the H3 N-tail influences H1 CTD condensation through direct protein-protein interaction, rather than alterations in linker DNA trajectory. These results support an emerging hypothesis wherein the H1 CTD serves as a nexus for signaling in the nucleosome.

    DOI: 10.1093/nar/gkaa949

    Web of Science

    Scopus

    PubMed

  21. L-DNA-tagged fluorescence: In situ hybridization for highly sensitive imaging of RNAs in single cells

    Motoyuki Ogata, Gosuke Hayashi, Anri Ichiu, Akimitsu Okamoto

    Organic and Biomolecular Chemistry   Vol. 18 ( 40 ) page: 8084 - 8088   2020.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Royal Society of Chemistry  

    We report an effective fluorescence in situ hybridization strategy, named l-DNA tagged FISH (LT-FISH), for highly sensitive RNA detection in fixed cultured cells. LT-FISH includes two-step hybridization processes with a l-d chimera oligonucleotide probe and a fluorescence-labeled PCR product tethering a l-DNA tag. The degree of fluorescence enhancement, depending on the length of PCR products, was up to 14-fold when the 606 bp product was used. Endogenous mRNA and miRNA in cancer cells were visualized by utilizing this l-DNA-mediated signal amplification technique.

    DOI: 10.1039/d0ob01635g

    Web of Science

    Scopus

    PubMed

  22. Toolbox for chemically synthesized histone proteins.

    Nakatsu K, Hayashi G, Okamoto A

    Current opinion in chemical biology   Vol. 58   page: 10 - 19   2020.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.cbpa.2020.04.016

    Web of Science

    Scopus

    PubMed

  23. Antibody-like proteins that capture and neutralize SARS-CoV-2

    Kondo, T; Iwatani, Y; Matsuoka, K; Fujino, T; Umemoto, S; Yokomaku, Y; Ishizaki, K; Kito, S; Sezaki, T; Hayashi, G; Murakami, H

    SCIENCE ADVANCES   Vol. 6 ( 42 )   2020.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Science Advances  

    To combat severe acute respiratory syndrome–related coronavirus 2 (SARS-CoV-2) and any unknown emerging pathogens in the future, the development of a rapid and effective method to generate high-affinity antibodies or antibody-like proteins is of critical importance. We here report high-speed in vitro selection of multiple high-affinity antibody-like proteins against various targets including the SARS-CoV-2 spike protein. The sequences of monobodies against the SARS-CoV-2 spike protein were successfully procured within only 4 days. Furthermore, the obtained monobody efficiently captured SARS-CoV-2 particles from the nasal swab samples of patients and exhibited a high neutralizing activity against SARS-CoV-2 infection (half-maximal inhibitory concentration, 0.5 nanomolar). High-speed in vitro selection of antibody-like proteins is a promising method for rapid development of a detection method for, and of a neutralizing protein against, a virus responsible for an ongoing, and possibly a future, pandemic.

    DOI: 10.1126/sciadv.abd3916

    Web of Science

    Scopus

    PubMed

  24. Fmoc-Compatible and C-terminal-Sequence-Independent Peptide Alkyl Thioester Formation Using Cysteinylprolyl Imide

    Nakatsu Koki, Yanase Masafumi, Hayashi Gosuke, Okamoto Akimitsu

    ORGANIC LETTERS   Vol. 22 ( 12 ) page: 4670 - 4674   2020.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/acs.orglett.0c01450

    Web of Science

    Scopus

    PubMed

  25. Novel Strategies of Peptide Ligation for Accelerating Chemical Protein Synthesis

    Hayashi Gosuke, Okamoto Akimitsu

    JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN   Vol. 78 ( 2 ) page: 130 - 139   2020.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Web of Science

  26. Novel Strategies of Peptide Ligation for Accelerating Chemical Protein Synthesis

    Hayashi Gosuke, Okamoto Akimitsu

    JOURNAL OF SYNTHETIC ORGANIC CHEMISTRY JAPAN   Vol. 78 ( 2 ) page: 130-139   2020.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

  27. タンパク質化学合成を活用した翻訳後修飾研究

    林 剛介

    ファルマシア   Vol. 56 ( 1 ) page: 46-50 - 50   2020

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:公益社団法人 日本薬学会  

    タンパク質化学合成法の発展により、多様な翻訳後修飾を持つタンパク質の作製が可能なってきた。本稿では、タンパク質化学合成法について解説するとともに、化学合成タンパク質を用いた翻訳後修飾研究について紹介する。特に、エピジェネティクス研究で中心的役割を果たすヒストンタンパク質の翻訳後修飾研究、また本特集のテーマであるユビキチン鎖やユビキチン化タンパク質を化学的に合成し、応用した研究例について紹介する。

    DOI: 10.14894/faruawpsj.56.1_46

    CiNii Research

  28. Novel Strategies of Peptide Ligation for Accelerating Chemical Protein Synthesis

    Hayashi Gosuke, Okamoto Akimitsu

    Journal of Synthetic Organic Chemistry, Japan   Vol. 78 ( 2 ) page: 130 - 139   2020

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:The Society of Synthetic Organic Chemistry, Japan  

    <p>Chemical protein synthesis (CPS) that consists of solid-phase peptide synthesis and peptide ligation generates not only naturally-occurring proteins with/without posttranslational modifications (PTMs), but also a variety of artificial proteins including unnatural amino-acids such as ᴅ-amino acids and fluorophore-labeled amino acids. This unique property offers new analytical methods for protein structure and interaction in terms of PTMs. In fact, we have chemically synthesized histone proteins with PTMs, which play an important role in regulation of gene expression in eukaryotic cells, and analyzed the effects of PTMs such as methylation, acetylation, and phosphorylation. However, current CPS is still in developing process and includes several issues to be solved such as difficult handling in hydrophobic protein synthesis and time-consuming multistep process for large protein synthesis. We have approached these issues by creating new strategies for peptide ligation. One-pot ligation of five peptide segments was demonstrated for the first time by utilizing multifunctionality of thiophenol compound in deprotection of allyloxycarbonyl group by palladium complex. New thioester precursors for native chemical ligation, which have potential to offer a novel two-way one-pot ligation, have also been developed recently. Furthermore, we have been developing a new strategy for simultaneous ligation of multiple peptides on DNA scaffold, which can connect peptides in highly diluted condition. This article also describes the background and current situation of CPS with our future perspectives.</p>

    DOI: 10.5059/yukigoseikyokaishi.78.130

    Scopus

    CiNii Research

  29. Chemical Synthesis of Cys-Containing Protein via Chemoselective Deprotection with Different Palladium Complexes

    Kamo Naoki, Hayashi Gosuke, Okamoto Akimitsu

    ORGANIC LETTERS   Vol. 21 ( 20 ) page: 8378 - 8382   2019.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/acs.orglett.9b03152

    Web of Science

    Scopus

    PubMed

  30. Cysteinylprolyl imide (CPI) peptide: a highly reactive and easily accessible crypto-thioester for chemical protein synthesis

    Yanase Masafumi, Nakatsu Koki, Cardos Charlane Joy, Konda Yoshiki, Hayashi Gosuke, Okamoto Akimitsu

    CHEMICAL SCIENCE   Vol. 10 ( 23 ) page: 5967 - 5975   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1039/c9sc00646j

    Web of Science

    Scopus

    PubMed

  31. Reaction design for highly efficient chemical protein synthesis

    Hayashi Gosuke, Okamoto Akimitsu

    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY   Vol. 257   page: .   2019.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

  32. Simultaneous and Traceless Ligation of Peptide Fragments on DNA Scaffold.

    Hayashi G, Yanase M, Nakatsuka Y, Okamoto A

    Biomacromolecules   Vol. 20 ( 3 ) page: 1246-1253 - 1253   2019.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/acs.biomac.8b01655

    PubMed

  33. Triple Function of 4-Mercaptophenylacetic Acid Promotes One-Pot Multiple Peptide Ligation.

    Kamo N, Hayashi G, Okamoto A

    Angewandte Chemie (International ed. in English)   Vol. 57 ( 50 ) page: 16533-16537 - 16537   2018.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/anie.201809765

    PubMed

  34. Chemistry-Driven Epigenetic Investigation of Histone and DNA Modifications.

    Sueoka T, Koyama K, Hayashi G, Okamoto A

    Chemical record (New York, N.Y.)   Vol. 18 ( 12 ) page: 1727-1744 - 1744   2018.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/tcr.201800040

    PubMed

  35. Efficient peptide ligation between allyl-protected Asp and Cys followed by palladium-mediated deprotection.

    Kamo N, Hayashi G, Okamoto A

    Chemical communications (Cambridge, England)   Vol. 54 ( 34 ) page: 4337-4340 - 4340   2018.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1039/c8cc01965g

    PubMed

  36. Hybridization-sensitive Fluorescent Oligonucleotide Probe Conjugated with Cell-penetrating Peptides for Enhanced Cellular Uptake

    Hayashi Gosuke, Tamai Makoto, Okamoto Akimitsu

    CHEMISTRY LETTERS   Vol. 46 ( 12 ) page: 1803-1806 - 1806   2017.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:The Chemical Society of Japan  

    <p>Intracellular delivery of oligonucleotides is essential not only for antisense- and RNAi-therapeutics, but also for live-cell RNA imaging with fluorescent nucleic acid probes. Herein, we developed oligonucleotide–peptide conjugates synthesized via Cu-catalyzed alkyne-azide cycloaddition between an exciton-controlled hybridization-sensitive oligonucleotide (ECHO) probe and linear or cyclic cell-penetrating peptides (CPPs). When lipofected to living HeLa cells, the CPP-conjugated ECHO probe showed enhanced cellular uptake efficiency compared with that of the unconjugated ECHO probe.</p>

    DOI: 10.1246/cl.170813

  37. Regulation of the Stability of the Histone H2A-H2B Dimer by H2A Tyr57 Phosphorylation.

    Sueoka T, Hayashi G, Okamoto A

    Biochemistry   Vol. 56 ( 36 ) page: 4767-4772 - 4772   2017.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/acs.biochem.7b00504

    PubMed

  38. Chemical synthesis of dual labeled proteins via differently protected alkynes enables intramolecular FRET analysis.

    Hayashi G, Kamo N, Okamoto A

    Chemical communications (Cambridge, England)   Vol. 53 ( 43 ) page: 5918-5921 - 5921   2017.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1039/c7cc02612a

    PubMed

  39. Chemically-activatable alkyne-tagged probe for imaging microdomains in lipid bilayer membranes.

    Yamaguchi S, Matsushita T, Izuta S, Katada S, Ura M, Ikeda T, Hayashi G, Suzuki Y, Kobayashi K, Tokunaga K, Ozeki Y, Okamoto A

    Scientific reports   Vol. 7   page: 41007   2017.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/srep41007

    PubMed

  40. Base-Resolution Analysis of 5-Hydroxymethylcytosine by One-Pot Bisulfite-Free Chemical Conversion with Peroxotungstate.

    Hayashi G, Koyama K, Shiota H, Kamio A, Umeda T, Nagae G, Aburatani H, Okamoto A

    Journal of the American Chemical Society   Vol. 138 ( 43 ) page: 14178-14181 - 14181   2016.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/jacs.6b06428

    PubMed

  41. In vitro and in cell analysis of chemically synthesized histone H2A with multiple modifications.

    Hayashi G, Sueoka T, Okamoto A

    Chemical communications (Cambridge, England)   Vol. 52 ( 28 ) page: 4999-5002 - 5002   2016.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1039/c5cc10555b

    PubMed

  42. Diazirine Photocrosslinking Recruits Activated FTO Demethylase Complexes for Specific N-6-methyladenosine Recognition Reviewed

    Hyun Seok Jeong, Gosuke Hayashi, Akimitsu Okamoto

    ACS CHEMICAL BIOLOGY   Vol. 10 ( 6 ) page: 1450 - 1455   2015.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    N-6-methyladenosine (m(6)A) is a prevalent modification of RNAs. m(6)A exists in mRNA and plays an important role in RNA biological pathways and in RNA epigenetic regulation, We applied diazirine photocrosslinking to the event of m(6)A recognition mediated by the fat mass and obesity associated (FTO) demethylase. A highly photoreactive diazirine adjacent to m(6)A on the RNA successfully recruited activated FTO complexes with an m(6)A preference. The process of recognition of m(6)A via FTO using diazirine photocrosslinking was controlled by the alpha-ketoglutarate (alpha-KG) cosubstrate and the Peal). cofactor, which are involved in m(6)A oxidative demethylation. In addition, FTO bound to ssRNAs prior to the m(6)A recognition process. Diazirine photocrosslinking contributes to increasing the chances of capturing activated FTO complexes with specific m(6)A recognition and provides new insights into the dynamic FTO oxidative demethylation process.

    DOI: 10.1021/cb5010096

    Web of Science

    PubMed

  43. 2-Oxazoline formation for selective chemical labeling of 5-hydroxylysine.

    Hayashi G, Sakamoto R, Okamoto A

    Chemistry, an Asian journal   Vol. 10 ( 5 ) page: 1138-41 - 41   2015.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/asia.201500172

    PubMed

  44. Hybridization-sensitive fluorescent oligonucleotide probe conjugated with a bulky module for compartment-specific mRNA monitoring in a living cell.

    Hayashi G, Yanase M, Takeda K, Sakakibara D, Sakamoto R, Wang DO, Okamoto A

    Bioconjugate chemistry   Vol. 26 ( 3 ) page: 412-7 - 7   2015.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/acs.bioconjchem.5b00090

    PubMed

  45. Middle-Down and Chemical Proteomic Approaches to Reveal Histone H4 Modification Dynamics in Cell Cycle: Label-Free Semi-Quantification of Histone Tail Peptide Modifications Including Phosphorylation and Highly Sensitive Capture of Histone PTM Binding Proteins Using Photo-Reactive Crosslinkers.

    Yamamoto K, Chikaoka Y, Hayashi G, Sakamoto R, Yamamoto R, Sugiyama A, Kodama T, Okamoto A, Kawamura T

    Mass spectrometry (Tokyo, Japan)   Vol. 4 ( 1 ) page: A0039 - A0039   2015

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:The Mass Spectrometry Society of Japan  

    Mass spectrometric proteomics is an effective approach for identifying and quantifying histone post-translational modifications (PTMs) and their binding proteins, especially in the cases of methylation and acetylation. However, another vital PTM, phosphorylation, tends to be poorly quantified because it is easily lost and inefficiently ionized. In addition, PTM binding proteins for phosphorylation are sometimes resistant to identification because of their variable binding affinities. Here, we present our efforts to improve the sensitivity of detection of histone H4 tail peptide phosphorylated at serine 1 (H4S1ph) and our successful identification of an H4S1ph binder candidate by means of a chemical proteomics approach.Our nanoLC-MS/MS system permitted semi-quantitative label-free analysis of histone H4 PTM dynamics of cell cycle-synchronized HeLa S3 cells, including phosphorylation, methylation, and acetylation. We show that H4S1ph abundance on nascent histone H4 unmethylated at lysine 20 (H4K20me0) peaks from late S-phase to M-phase. We also attempted to characterize effects of phosphorylation at H4S1 on protein–protein interactions. Specially synthesized photoaffinity bait peptides specifically captured 14-3-3 proteins as novel H4S1ph binding partners, whose interaction was otherwise undetectable by conventional peptide pull-down experiments.This is the first report that analyzes dynamics of PTM pattern on the whole histone H4 tail during cell cycle and enables the identification of PTM binders with low affinities using high-resolution mass spectrometry and photo-affinity bait peptides.

    DOI: 10.5702/massspectrometry.A0039

    PubMed

  46. An orthogonal ribosome-tRNA pair via engineering of the peptidyl transferase center.

    Terasaka N, Hayashi G, Katoh T, Suga H

    Nature chemical biology   Vol. 10 ( 7 ) page: 555-7 - 557   2014.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nature Publishing Group  

    DOI: 10.1038/nchembio.1549

    PubMed

    CiNii Books

  47. Development of photoswitchable RNA aptamer-ligand complexes.

    Hayashi G, Nakatani K

    Methods in molecular biology (Clifton, N.J.)   Vol. 1111   page: 29-40 - 40   2014

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1007/978-1-62703-755-6_3

    PubMed

  48. Probe design for the effective fluorescence imaging of intracellular RNA.

    Hayashi G, Okamoto A

    Chemical record (New York, N.Y.)   Vol. 13 ( 2 ) page: 209-17 - 17   2013.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/tcr.201200026

    PubMed

  49. A nucleic acid probe labeled with desmethyl thiazole orange: a new type of hybridization-sensitive fluorescent oligonucleotide for live-cell RNA imaging.

    Okamoto A, Sugizaki K, Yuki M, Yanagisawa H, Ikeda S, Sueoka T, Hayashi G, Wang DO

    Organic & biomolecular chemistry   Vol. 11 ( 2 ) page: 362-71 - 71   2013.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1039/c2ob26707a

    PubMed

  50. Activation of prokaryotic translation by antisense oligonucleotides binding to coding region of mRNA.

    Hayashi G, Hong C, Hagihara M, Nakatani K

    Biochemical and biophysical research communications   Vol. 429 ( 1-2 ) page: 105-10 - 10   2012.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.bbrc.2012.10.072

    PubMed

  51. The RNA origin of transfer RNA aminoacylation and beyond.

    Suga H, Hayashi G, Terasaka N

    Philosophical transactions of the Royal Society of London. Series B, Biological sciences   Vol. 366 ( 1580 ) page: 2959-64 - 64   2011.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1098/rstb.2011.0137

    PubMed

  52. [Ribosomal synthesis of nonstandard cyclic peptides and its application to drug discovery].

    Hayashi G, Ohshiro Y, Suga H

    Seikagaku. The Journal of Japanese Biochemical Society   Vol. 82 ( 6 ) page: 505-14   2010.6

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)  

  53. [Ribosomal synthesis of nonstandard cyclic peptides and its application to drug discovery].

    Hayashi G, Ohshiro Y, Suga H

    Seikagaku. The Journal of Japanese Biochemical Society   Vol. 82 ( 6 ) page: 505 - 14   2010.6

     More details

    Publishing type:Research paper (scientific journal)  

    PubMed

  54. Ribosome evolution for two artificial amino acids in E. coli. International journal

    Hayashi G, Goto Y, Suga H

    Chemistry & biology   Vol. 17 ( 4 ) page: 320-1 - 1   2010.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.chembiol.2010.04.005

    PubMed

  55. RNA aptamers that reversibly bind photoresponsive azobenzene-containing peptides.

    Hayashi G, Hagihara M, Nakatani K

    Chemistry (Weinheim an der Bergstrasse, Germany)   Vol. 15 ( 2 ) page: 424-32 - 32   2009

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/chem.200800936

    PubMed

  56. Genotyping by allele-specific L-DNA-tagged PCR.

    Hayashi G, Hagihara M, Nakatani K

    Journal of biotechnology   Vol. 135 ( 2 ) page: 157-60 - 60   2008.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.jbiotec.2008.03.011

    PubMed

  57. RNA aptamers that reversibly bind to photoresponsive peptide.

    Hayashi G, Hagihara M, Nakatani K

    Nucleic acids symposium series (2004)   ( 52 ) page: 703-4 - 4   2008

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nass/nrn355

    PubMed

  58. Photoregulation of a peptide-RNA interaction on a gold surface.

    Hayashi G, Hagihara M, Dohno C, Nakatani K

    Journal of the American Chemical Society   Vol. 129 ( 28 ) page: 8678-9 - 9   2007.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1021/ja071298x

    PubMed

  59. Detection of L-DNA-tagged PCR products by surface plasmon resonance imaging.

    Hayashi G, Hagihara M, Kobori A, Nakatani K

    Chembiochem : a European journal of chemical biology   Vol. 8 ( 2 ) page: 169-71 - 71   2007.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/cbic.200600477

    PubMed

  60. Reversible regulation of binding between a photoresponsive peptide and its RNA aptamer.

    Hayashi G, Hagihara M, Dohno C, Nakatani K

    Nucleic acids symposium series (2004)   ( 51 ) page: 93-4 - 4   2007

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nass/nrm047

    PubMed

  61. N,N'-Bis(3-aminopropyl)-2,7-diamino-1,8-naphthyridine stabilized a single pyrimidine bulge in duplex DNA.

    Suda H, Kobori A, Zhang J, Hayashi G, Nakatani K

    Bioorganic & medicinal chemistry   Vol. 13 ( 14 ) page: 4507-12 - 12   2005.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.bmc.2005.04.035

    PubMed

  62. Small-molecule ligand induces nucleotide flipping in (CAG)n trinucleotide repeats.

    Nakatani K, Hagihara S, Goto Y, Kobori A, Hagihara M, Hayashi G, Kyo M, Nomura M, Mishima M, Kojima C

    Nature chemical biology   Vol. 1 ( 1 ) page: 39-43 - 43   2005.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/nchembio708

    PubMed

  63. Application of L-DNA as a molecular tag.

    Hayashi G, Hagihara M, Nakatani K

    Nucleic acids symposium series (2004)   ( 49 ) page: 261-2 - 2   2005

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nass/49.1.261

    PubMed

  64. Solution structure of a small-molecular ligand complexed with CAG trinucleotide repeat DNA.

    Nakatani K, Hagihara S, Goto Y, Kobori A, Hagihara M, Hayashi G, Kyo M, Nomura M, Mishima M, Kojima C

    Nucleic acids symposium series (2004)   ( 49 ) page: 49-50 - 50   2005

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nass/49.1.49

    PubMed

  65. Detection of guanine-adenine mismatches by surface plasmon resonance sensor carrying naphthyridine-azaquinolone hybrid on the surface. International journal

    Hagihara S, Kumasawa H, Goto Y, Hayashi G, Kobori A, Saito I, Nakatani K

    Nucleic acids research   Vol. 32 ( 1 ) page: 278-86 - 86   2004

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We have discovered a new molecule naphthyridine-azaquinolone hybrid (Npt-Azq) that strongly stabilized the guanine-adenine (G-A) mismatch in duplex DNA. In the presence of Npt-Azq, the melting temperature (T(m)) of 5'-d(CTA ACG GAA TG)-3'/3'-d(GAT TGA CTT AC)-5' containing a single G-A mismatch increased by 15.4 degrees C, whereas fully matched duplex increased its T(m) only by 2.2 degrees C. Npt-Azq was immobilized on the sensor surface for the surface plasmon resonance (SPR) assay to examine SPR detection of duplexes containing a G-A mismatch. Distinct SPR signals were observed when 27mer DNA containing a G-A mismatch was analyzed by the Npt-Azq immobilized sensor surfaces, whereas the signal of the fully matched duplex was approximately 6-fold weaker in intensity. The SPR signals for the G-A mismatch were proportional to the concentration of DNA in a range up to 1 microM, confirming that the SPR signal is in fact due to the binding of the G-A mismatch to Npt-Azq immobilized on the surface. Examination of all 16 G-A mismatches regarding the flanking sequence revealed that the sensor surface reported here is applicable to eight flanking sequences, covering 50% of all possible G-A mismatches.

    DOI: 10.1093/nar/gkh171

    PubMed

  66. SPR fingerprinting of mismatched base pair.

    Kobori A, Peng T, Hayashi G, Nakatani K

    Nucleic acids symposium series (2004)   ( 48 ) page: 129-30 - 30   2004

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nass/48.1.129

    PubMed

▼display all

MISC 28

  1. Reaction design for highly efficient chemical protein synthesis

    Hayashi Gosuke, Okamoto Akimitsu

    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY   Vol. 257   page: .   2019.3

     More details

    Language:English  

    Web of Science

  2. Hybridization-sensitive Fluorescent Oligonucleotide Probe Conjugated with Cell-penetrating Peptides for Enhanced Cellular Uptake

    Hayashi Gosuke, Tamai Makoto, Okamoto Akimitsu

    CHEMISTRY LETTERS   Vol. 46 ( 12 ) page: 1803-1806   2017.12

     More details

    Language:English  

    DOI: 10.1246/cl.170813

    Web of Science

  3. Base-Resolution Analysis of 5-Hydroxymethylcytosine by One-Pot Bisulfite-Free Chemical Conversion with Peroxotungstate.

    Hayashi G, Koyama K, Shiota H, Kamio A, Umeda T, Nagae G, Aburatani H, Okamoto A

    Journal of the American Chemical Society   Vol. 138 ( 43 ) page: 14178-14181   2016.11

     More details

    Language:English  

    DOI: 10.1021/jacs.6b06428

    PubMed

  4. In vitro and in cell analysis of chemically synthesized histone H2A with multiple modifications.

    Hayashi G, Sueoka T, Okamoto A

    Chemical communications (Cambridge, England)   Vol. 52 ( 28 ) page: 4999-5002   2016.4

     More details

    Language:English  

    DOI: 10.1039/c5cc10555b

    PubMed

  5. Diazirine photocrosslinking recruits activated FTO demethylase complexes for specific N(6)-methyladenosine recognition.

    Jeong HS, Hayashi G, Okamoto A

    ACS chemical biology   Vol. 10 ( 6 ) page: 1450-5   2015.6

     More details

    Language:English  

    DOI: 10.1021/cb5010096

    PubMed

  6. 2-Oxazoline formation for selective chemical labeling of 5-hydroxylysine.

    Hayashi G, Sakamoto R, Okamoto A

    Chemistry, an Asian journal   Vol. 10 ( 5 ) page: 1138-41   2015.5

     More details

    Language:English  

    DOI: 10.1002/asia.201500172

    PubMed

  7. Hybridization-sensitive fluorescent oligonucleotide probe conjugated with a bulky module for compartment-specific mRNA monitoring in a living cell.

    Hayashi G, Yanase M, Takeda K, Sakakibara D, Sakamoto R, Wang DO, Okamoto A

    Bioconjugate chemistry   Vol. 26 ( 3 ) page: 412-7   2015.3

     More details

    Language:English  

    DOI: 10.1021/acs.bioconjchem.5b00090

    PubMed

  8. Middle-Down and Chemical Proteomic Approaches to Reveal Histone H4 Modification Dynamics in Cell Cycle: Label-Free Semi-Quantification of Histone Tail Peptide Modifications Including Phosphorylation and Highly Sensitive Capture of Histone PTM Binding Proteins Using Photo-Reactive Crosslinkers.

    Yamamoto K, Chikaoka Y, Hayashi G, Sakamoto R, Yamamoto R, Sugiyama A, Kodama T, Okamoto A, Kawamura T

    Mass spectrometry (Tokyo, Japan)   Vol. 4 ( 1 ) page: A0039   2015

     More details

    Language:English  

    DOI: 10.5702/massspectrometry.A0039

    PubMed

  9. An orthogonal ribosome-tRNA pair via engineering of the peptidyl transferase center.

    Terasaka N, Hayashi G, Katoh T, Suga H

    Nature chemical biology   Vol. 10 ( 7 ) page: 555-7   2014.7

     More details

    Language:English  

    DOI: 10.1038/nchembio.1549

    PubMed

  10. Development of photoswitchable RNA aptamer-ligand complexes.

    Hayashi G, Nakatani K

    Methods in molecular biology (Clifton, N.J.)   Vol. 1111   page: 29-40   2014

     More details

    Language:English  

    DOI: 10.1007/978-1-62703-755-6_3

    PubMed

  11. Probe design for the effective fluorescence imaging of intracellular RNA.

    Hayashi G, Okamoto A

    Chemical record (New York, N.Y.)   Vol. 13 ( 2 ) page: 209-17   2013.4

     More details

    Language:English  

    DOI: 10.1002/tcr.201200026

    PubMed

  12. A nucleic acid probe labeled with desmethyl thiazole orange: a new type of hybridization-sensitive fluorescent oligonucleotide for live-cell RNA imaging.

    Okamoto A, Sugizaki K, Yuki M, Yanagisawa H, Ikeda S, Sueoka T, Hayashi G, Wang DO

    Organic & biomolecular chemistry   Vol. 11 ( 2 ) page: 362-71   2013.1

     More details

    Language:English  

    DOI: 10.1039/c2ob26707a

    PubMed

  13. Activation of prokaryotic translation by antisense oligonucleotides binding to coding region of mRNA.

    Hayashi G, Hong C, Hagihara M, Nakatani K

    Biochemical and biophysical research communications   Vol. 429 ( 1-2 ) page: 105-10   2012.12

     More details

    Language:English  

    DOI: 10.1016/j.bbrc.2012.10.072

    PubMed

  14. The RNA origin of transfer RNA aminoacylation and beyond.

    Suga H, Hayashi G, Terasaka N

    Philosophical transactions of the Royal Society of London. Series B, Biological sciences   Vol. 366 ( 1580 ) page: 2959-64   2011.10

     More details

    Language:English  

    DOI: 10.1098/rstb.2011.0137

    PubMed

  15. [Ribosomal synthesis of nonstandard cyclic peptides and its application to drug discovery].

    Hayashi G, Ohshiro Y, Suga H

    Seikagaku. The Journal of Japanese Biochemical Society   Vol. 82 ( 6 ) page: 505-14   2010.6

     More details

    Language:Japanese  

    PubMed

  16. Ribosome evolution for two artificial amino acids in E. coli.

    Hayashi G, Goto Y, Suga H

    Chemistry & biology   Vol. 17 ( 4 ) page: 320-1   2010.4

     More details

    Language:English  

    DOI: 10.1016/j.chembiol.2010.04.005

    PubMed

  17. RNA aptamers that reversibly bind photoresponsive azobenzene-containing peptides.

    Hayashi G, Hagihara M, Nakatani K

    Chemistry (Weinheim an der Bergstrasse, Germany)   Vol. 15 ( 2 ) page: 424-32   2009

     More details

    Language:English  

    DOI: 10.1002/chem.200800936

    PubMed

  18. Genotyping by allele-specific L-DNA-tagged PCR.

    Hayashi G, Hagihara M, Nakatani K

    Journal of biotechnology   Vol. 135 ( 2 ) page: 157-60   2008.6

     More details

    Language:English  

    DOI: 10.1016/j.jbiotec.2008.03.011

    PubMed

  19. RNA aptamers that reversibly bind to photoresponsive peptide.

    Hayashi G, Hagihara M, Nakatani K

    Nucleic acids symposium series (2004)   ( 52 ) page: 703-4   2008

     More details

    Language:English  

    DOI: 10.1093/nass/nrn355

    PubMed

  20. Photoregulation of a peptide-RNA interaction on a gold surface.

    Hayashi G, Hagihara M, Dohno C, Nakatani K

    Journal of the American Chemical Society   Vol. 129 ( 28 ) page: 8678-9   2007.7

     More details

    Language:English  

    DOI: 10.1021/ja071298x

    PubMed

  21. Detection of L-DNA-tagged PCR products by surface plasmon resonance imaging.

    Hayashi G, Hagihara M, Kobori A, Nakatani K

    Chembiochem : a European journal of chemical biology   Vol. 8 ( 2 ) page: 169-71   2007.1

     More details

    Language:English  

    DOI: 10.1002/cbic.200600477

    PubMed

  22. Reversible regulation of binding between a photoresponsive peptide and its RNA aptamer.

    Hayashi G, Hagihara M, Dohno C, Nakatani K

    Nucleic acids symposium series (2004)   ( 51 ) page: 93-4   2007

     More details

    Language:English  

    DOI: 10.1093/nass/nrm047

    PubMed

  23. N,N'-Bis(3-aminopropyl)-2,7-diamino-1,8-naphthyridine stabilized a single pyrimidine bulge in duplex DNA.

    Suda H, Kobori A, Zhang J, Hayashi G, Nakatani K

    Bioorganic & medicinal chemistry   Vol. 13 ( 14 ) page: 4507-12   2005.7

     More details

    Language:English  

    DOI: 10.1016/j.bmc.2005.04.035

    PubMed

  24. Small-molecule ligand induces nucleotide flipping in (CAG)n trinucleotide repeats.

    Nakatani K, Hagihara S, Goto Y, Kobori A, Hagihara M, Hayashi G, Kyo M, Nomura M, Mishima M, Kojima C

    Nature chemical biology   Vol. 1 ( 1 ) page: 39-43   2005.6

     More details

    Language:English  

    DOI: 10.1038/nchembio708

    PubMed

  25. Application of L-DNA as a molecular tag.

    Hayashi G, Hagihara M, Nakatani K

    Nucleic acids symposium series (2004)   ( 49 ) page: 261-2   2005

     More details

    Language:English  

    DOI: 10.1093/nass/49.1.261

    PubMed

  26. Solution structure of a small-molecular ligand complexed with CAG trinucleotide repeat DNA.

    Nakatani K, Hagihara S, Goto Y, Kobori A, Hagihara M, Hayashi G, Kyo M, Nomura M, Mishima M, Kojima C

    Nucleic acids symposium series (2004)   ( 49 ) page: 49-50   2005

     More details

    Language:English  

    DOI: 10.1093/nass/49.1.49

    PubMed

  27. SPR fingerprinting of mismatched base pair.

    Kobori A, Peng T, Hayashi G, Nakatani K

    Nucleic acids symposium series (2004)   ( 48 ) page: 129-30   2004

     More details

    Language:English  

    DOI: 10.1093/nass/48.1.129

    PubMed

  28. Detection of guanine-adenine mismatches by surface plasmon resonance sensor carrying naphthyridine-azaquinolone hybrid on the surface.

    Hagihara S, Kumasawa H, Goto Y, Hayashi G, Kobori A, Saito I, Nakatani K

    Nucleic acids research   Vol. 32 ( 1 ) page: 278-86   2004

     More details

    Language:English  

    DOI: 10.1093/nar/gkh171

    PubMed

▼display all

Research Project for Joint Research, Competitive Funding, etc. 2

  1. Sタンパク質結合人工抗体を用いたコロナウィルス濃縮および抗原検査法の開発

    2020.7 - 2021.6

    出資金による受託研究 

  2. 有機化学を基盤としたエピゲノム修飾ヌク レオソーム再構成技術の確立

    2019.10 - 2023.3

    特色ある大学教育支援プログラム 

      More details

    Grant type:Competitive

KAKENHI (Grants-in-Aid for Scientific Research) 20

  1. 膜脂質のユビキチン化とユビキチン様タンパク質修飾によるオルガネラ機能制御

    Grant number:24K01971  2024.4 - 2027.3

    科学研究費助成事業  基盤研究(B)

    坂巻 純一, 林 剛介

      More details

    Authorship:Coinvestigator(s) 

    オルガネラ上では、シグナル伝達、生化学反応、物質輸送などが行われている。これまで、膜タンパク質の修飾に着目したオルガネラ機能制御の研究が数多くなされてきたが、膜を構成する脂質の変化に着目した解析はほとんどない。研究代表者は、オルガネラ膜の脂質がユビキチン化されるという新しい膜脂質修飾を発見した。しかし、膜脂質のユビキチン化の分子機構や生物学的意義は不明である。そこで、本研究では、膜脂質のユビキチン化とユビキチン様タンパク質修飾の解析に基軸を置き、修飾の分子機構とその破綻が細胞機能に及ぼす障害の解明を目標とする。膜脂質のユビキチン化修飾という新しい視点から、オルガネラの機能制御の理解を目指す。

  2. タンパク質寿命の解析・制御研究をブーストする人工タンパク質創成技術

    Grant number:24H01893  2024.4 - 2026.3

    科学研究費助成事業  学術変革領域研究(A)

    林 剛介

      More details

    Authorship:Principal investigator 

    Grant amount:\9360000 ( Direct Cost: \7200000 、 Indirect Cost:\2160000 )

    本研究では、タンパク質寿命を決定する分子メカニズムの解明、およびタンパク質寿命の制御技術確立を見据え、有機化学および進化分子工学による人工タンパク質創成技術の開発およびシン(進/深)化を行う。具体的には、「タンパク質化学合成法」の新手法を開発し、ユビキチン化などタンパク質寿命の決定に関わる翻訳後修飾が導入されたタンパク質を合成する。さらに「in vitroセレクション法」によって、ユビキチンリガーゼや寿命制御の標的タンパク質に結合する人工抗体を取得し、人工抗体デグレーダーを創成する。また、これら両技術を融合し、化学合成で調製したユビキチン化タンパク質に結合する人工抗体を取得する。

  3. Development of single-molecule peptide sequencing method

    Grant number:23H05456  2023.4 - 2028.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (S)

      More details

    Authorship:Coinvestigator(s) 

  4. International collaboration for the development of new molecular-targeted antimicrobial agents

    Grant number:22KK0128  2022.10 - 2026.3

    科学研究費助成事業  国際共同研究加速基金(国際共同研究強化(B))

    Kim Minsoo, Raudzus Fabian, 林 剛介, 西出 旭

      More details

    Authorship:Coinvestigator(s) 

    抗生物質を手にした現代においても、細菌感染症は人類にとって大きな脅威である。抗菌剤には多剤耐性菌の出現など様々な問題があり、現在の抗菌剤とは作用機序の異なる治療薬の開発が必要である。国際共同研究により、細菌感染症に対する治療薬の不足という世界的な課題に取り込むことが本研究の目的である。
    本研究では、病原細菌が宿主細胞に分泌する病原因子に着目し、 (1)標的とする病原因子とその宿主蛋白質との結合様式を解明する、 (2)海外共同研究者がもつ最新の質量分析技術を活用して、当該病原因子による宿主蛋白質の制御メカニズムを明らかにする、(3)その制御メカニズムを狙った新規抗菌剤を開発することを目指す。

  5. Ubiquitin Chemo-technology by Chemical Protein Synthesis and Molecular Evolution

    Grant number:21H00278  2021.4 - 2023.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

      More details

    Authorship:Principal investigator 

    Grant amount:\7280000 ( Direct Cost: \5600000 、 Indirect Cost:\1680000 )

  6. Sタンパク質結合人工抗体を用いたコロナウィルス濃縮および抗原検査法の開発

    2020.7 - 2021.6

    出資金による受託研究 

      More details

    Grant type:Competitive

  7. 生体分子夾雑系で機能するD体人工抗体の開発

    2020.4 - 2022.3

    科学研究費補助金 

      More details

    Authorship:Principal investigator 

  8. 生体分子夾雑系で機能するD体人工抗体の開発

    2020.4 - 2022.3

    科学研究費助成事業  新学術領域研究(研究領域提案型)

      More details

    Grant type:Competitive

  9. Mirror-image Artificial Antibody Working in Multimolecular Crowding Biosystems

    Grant number:20H04704  2020.4 - 2022.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

      More details

    Authorship:Principal investigator 

    Grant amount:\4940000 ( Direct Cost: \3800000 、 Indirect Cost:\1140000 )

  10. 有機化学を基盤としたエピゲノム修飾ヌク レオソーム再構成技術の確立

    2019.10 - 2023.3

    文部科学省  戦略的創造研究推進事業 

      More details

    Grant type:Competitive

  11. Efficient Chemical Synthesis of Polyubiquitin and Ubiquitinated Proteins

    Grant number:19H05287  2019.4 - 2021.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

      More details

    Authorship:Principal investigator 

    Grant amount:\5070000 ( Direct Cost: \3900000 、 Indirect Cost:\1170000 )

  12. DNAを足場としたペプチド断片の同時連結反応によるタンパク質の効率的化学合成

    2018.4 - 2021.3

    科学研究費補助金  基盤研究(C)

      More details

    Authorship:Principal investigator 

  13. DNAを足場としたペプチド断片の同時連結反応によるタンパク質の効率的化学合成

    2018.4 - 2021.3

    日本学術振興会  科学研究費助成事業  基盤研究(C)

      More details

    Grant type:Competitive

  14. Chemical synthesis of protein via simultaneous ligation of multiple peptide segments on DNA scaffold

    Grant number:18K05313  2018.4 - 2021.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Hayashi Gosuke

      More details

    Authorship:Principal investigator 

    Grant amount:\4420000 ( Direct Cost: \3400000 、 Indirect Cost:\1020000 )

    We have developed a novel peptide ligation strategy harnessing the two intrinsic characteristics of oligodeoxynucleotides (ODNs), i.e., their hydrophilicity and hybridization ability, which allowed an increase in the water solubility of peptides and the reaction kinetics due to the proximity effect, respectively. Peptide&#8211;ODN conjugates cleavable to regenerate native peptide sequences were synthesized using novel lysine derivatives containing conjugation handles and photolabile linkers, via solid-phase peptide synthesis and subsequent conjugation to 15-mer ODNs. Two complementary conjugates were applied to carbodiimide-mediated peptide ligation on a DNA scaffold and the subsequent DNA removal was conducted by photoirradiation in a traceless fashion.

  15. 分解反応の遷移状態構造に立脚した新型核酸医薬を志向した核酸酵素の創製

    2015.7 - 2018.3

    科学研究費補助金  基盤研究(B)

      More details

    Authorship:Coinvestigator(s) 

  16. 分解反応の遷移状態構造に立脚した新型核酸医薬を志向した核酸酵素の創製

    2015.7 - 2018.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

      More details

    Grant type:Competitive

  17. Development of nucleic acid enzymes for new nucleic acid medicines based on the transition state in molecular degradation

    Grant number:15KT0057  2015.7 - 2018.3

    Okamoto Akimitsu

      More details

    Authorship:Coinvestigator(s) 

    The solid-state fermentation (SSF) bioprocesses are performed in complex solid-rich systems that present significant challenges for effective monitoring of bacterial population dynamics. We have developed an efficient chemical system that allows quantification of bacteria population by fluorescence-based analysis. The key component in the system is the exciton-controlled fluorescent RNA aptamer, which was covalently conjugated to two thiazole orange moieties and serves as a competitor of bacterial ribosome. The intensity of fluorescence from such a ribosome-sensing system was controlled by the excitonic interaction between dyes in the RNA aptamer and it increased drastically in the presence of Escherichia coli. This innovative fluorescence-based competition system is valuable for quantification without any extraction of bacterial nucleic acids, and provides the simplest and most feasible way to optimize SSF bioprocesses.

  18. ヒストンコード研究の基盤となる多様な修飾を有するヌクレオソーム構築法の開発と応用

    2013.4 - 2015.3

    科学研究費補助金  若手研究(B)

      More details

    Authorship:Principal investigator 

  19. ヒストンコード研究の基盤となる多様な修飾を有するヌクレオソーム構築法の開発と応用

    2013.4 - 2015.3

    日本学術振興会  科学研究費助成事業  若手研究(B)

      More details

    Grant type:Competitive

  20. Development of a platform for histone-code research by setting nucleosomes with various modifications

    Grant number:25870186  2013.4 - 2015.3

    HAYASHI Gosuke

      More details

    Authorship:Principal investigator 

    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

    We developed a chemical platform for approaching “histone code hypothesis”, that is one of the most important issues in epigenetics research. Total chemical synthesis of core histone H2A, H2B, and linker histone H1, all of which play essential roles in gene regulation and chromatin integrity, have been achieved by solid-phase peptide synthesis (SPPS) and native chemical ligation (NCL). The chemically-synthesized histones showed comparable ability to form nucleosome and chromatosome in vitro. Furthermore, we introduced dye-labelled H2A into HeLa cells and observed that the synthetic histone protein successfully localized into nucleus.

▼display all

 

Teaching Experience (On-campus) 12

  1. 生体分子応用化学

    2020

  2. 分析化学3

    2020

  3. 化学生命工学実験3

    2020

  4. 分子生命化学基礎論

    2020

  5. 化学実験

    2020

  6. 化学生命工学実験1

    2020

  7. 分析化学2

    2020

  8. G30 Core-Biochemistry

    2019

  9. 化学生命工学実験3

    2019

  10. 分析化学3

    2019

  11. 化学生命工学実験1

    2019

  12. 分析化学2

    2019

▼display all

Teaching Experience (Off-campus) 13

  1. G30 Core-Biochemistry

    Nagoya University)

  2. G30 Core-Biochemistry

    Nagoya University)

  3. 分子生命化学基礎論

    Nagoya University)

  4. 生体分子応用化学

    Nagoya University)

  5. 化学生命工学実験3

    Nagoya University)

  6. 化学生命工学実験3

    Nagoya University)

  7. 化学生命工学実験1

    Nagoya University)

  8. 化学生命工学実験1

    Nagoya University)

  9. 化学実験

    Nagoya University)

  10. 分析化学3

    Nagoya University)

  11. 分析化学3

    Nagoya University)

  12. 分析化学2

    Nagoya University)

  13. 分析化学2

    Nagoya University)

▼display all