2007 Fiscal Year Final Research Report Summary
Chemical Expansion of the Central Dogma towards Synthetic Microorganisms
| Project/Area Number |
15101008
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| Research Category |
Grant-in-Aid for Scientific Research (S)
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| Allocation Type | Single-year Grants |
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| Research Field |
Living organism molecular science
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| Research Institution | Okayama University |
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Principal Investigator |
SISIDO Masahiko Okayama University, Graduate School of Natura Science, Professor (60026268)
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| Co-Investigator(Kenkyū-buntansha) |
OHTSUKI Takashi Okayama University, Graduate School of Natura Science, Associate Professor (80321735)
TAKI Masumi Okayama University, Graduate School of Natura Science, Assistant Professor (70362952)
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| Project Period (FY) |
2003 – 2007
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| Keywords | Nonnatural amino acid / In vitro protein synthesis / Aminoacylation / tRNA / Nonnatural mutant protein / In vivo protein synthesis / Synthetic microorganism |
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| Research Abstract |
This research project aims to expand molecules like amino acids, nucleic acids and proteins constitute the protein biosynthesizing system to synthesize novel type of proteins with artificial functions. The final goal is to create synthetic microorganisms that live with 21 type of amino acids. During the 5 year we obtained the following results. (1) Some nonnatural amino acids were successfully charged onto a specific tRNA by the use of a peptide nucleic acid (PNA) that is complementary to the tRNA. The PNA-assisted aminoacylation mold be carried out in an E.coli in vitro protein biosynthesizing system to create a protein incorporated with the nonnatural amino add. (2) tRNAs and an elongation faint Tu. (EF-Tu) were expanded to bring large-sized nonnatural amino adds into E.col ribosome and subsequently to a protein. (3) An orthogonal set of aminoacyl tRNAsynthetase (ARS) and tRNA was screened to bring some nonnatural amino acids into proteins in living mammalian cells. (4) Finally these expanded molecular systems were introduced into mammalian cells and the synthesis of proteins containing nonnatural amino acids was detected, although its quantity was very small.
In the last year, an attempt was made to apply these techniques to attach fluorescent amino acids to peptides and proteins. The fluorescently labeled peptides were screened to dimmer those that bind to cancer cells or to cancer-cell specific proteins.
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[Journal Article] FRET analysis of protein conformational change through position-specific incorporation of fluorescent amino acids2006
Author(s)
D., Kajihara, R., Abe, I., lijima, C., Komiyama, M., Sisido, T., Hohsaka
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Journal Title
Nature Methods 3
Pages: 923-929
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Synthesis of novel gem-difluorinatedcyclopropane hybrids : Applications for DNA cleavage agents switched by photo irradiation2006
Author(s)
K., Ninomiya, K., Tanimoto, N., Ishida, D., Horii, M., Sisido, T., Rob
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Journal Title
J. Fluorine Chem 127
Pages: 651-656
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Leucyl/Phenylalanyl-tRNA-protein transferase-mediated chemo-enzymatic coupling of N-terminal Arg/Lys units in post-translationally processed proteins with nonnatural amino acids2006
Author(s)
M., Taki, A., Kuno, S., Matoba, Y., Kobayashi, J., Futami, H., Murakami, H., Suga, K., Taira, T., Hasegawa, M., Sisido
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Journal Title
ChemBioChem 7
Pages: 1676-1679
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] An evolutionary "intermediate" state of mitochondrial translation system found in parasitic nematode, Trichinel la spp. : Co-evolution of tRNA and EF-Tu2006
Author(s)
M., Arita, T., Suematsu, A., Osanai, T., Inaba, H., Kamiya, K., Kita, M., Sisido, Yoh-ichi, Watanabe, T., Ohtsuki
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Journal Title
Nucleic Acids Res 34
Pages: 5291-5299
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Position-Specific Incorporation of a Highly Photodurable and Blue-Laser Excitable Fluorescent Amino Acid into Proteins for Fluorescence Sensing2005
Author(s)
H., Hamada, N., Kameshima, A., Szymanska, K., Wegner, L., Lankiewicz, H., Shinohara, M., Taki, M., Sisido
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Journal Title
Bioorg. Med. Chem 13(10)
Pages: 3379-3384
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Incorporation of Anthraquinonyl Group into X-Cro Repressor Protein for Strand- and Position-Specific Photocleavage of Double-Stranded DNA2004
Author(s)
H., Sasaki, K., Ikeda, M., Suzuki, K., Ninomiya, M., Sisido
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Journal Title
Biopolymers, Peptide Science 76(1)
Pages: 21-26
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Position-Specific Incorporation of Dansylated Nonnatural Amino Acids into Streptavidin by using a four-base codon2004
Author(s)
T., Hohsaka, N., Muranaka, C., Komiyama, K., Matsui, S., Takaura, R., Abe, H., Murakami, M., Sisido
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Journal Title
FEBS Lett 560
Pages: 173-177
Description
「研究成果報告書概要(欧文)」より
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[Presentation] RNA isolation using biotinylated PNAs2007
Author(s)
T. Ohtsuki, T. Fujimoto, M. Kamimukai, M. Kitamatsu, M. Sisido
Organizer
The 2nd International Workshop on Approaches to Single Cell Analysis
Place of Presentation
Tokyo
Year and Date
20070906-07
Description
「研究成果報告書概要(和文)」より
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[Presentation] tRNA Aminoacylation Using Peptide Nucleic Acid as a Carrier of Amino Acid Esters2007
Author(s)
M., Sisido, M., Nishimura, T., Minohata, K., Ninomiya, M., Kitamatsu
Organizer
The 9th Chinese International Peptide Symposium International Convention Center, (Invited lecture)
Place of Presentation
Shanghai, China
Year and Date
2007-07-05
Description
「研究成果報告書概要(欧文)」より
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