Site-specific incorporation of non-natural amino acids into proteins in living cells
Project/Area Number |
19380195
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Applied molecular and cellular biology
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Research Institution | The Institute of Physical and Chemical Research |
Principal Investigator |
SAKAMOTO Kensaku The Institute of Physical and Chemical Research, 拡張遺伝暗号システム研究チーム, チームリーダー (50240685)
|
Project Period (FY) |
2007 – 2009
|
Project Status |
Completed (Fiscal Year 2009)
|
Budget Amount *help |
¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000)
Fiscal Year 2009: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2008: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2007: ¥11,570,000 (Direct Cost: ¥8,900,000、Indirect Cost: ¥2,670,000)
|
Keywords | 非天然型アミノ酸 / 遺伝暗号 / タンパク質生合成 / 光クロスリンク / 翻訳後修飾 / 蛍光修飾 / 昆虫細胞 / 蛍光標識 / アセチルリジン / ピロリジルtRNA合成酵素 / 動物細胞 / アミノアシルtRNA合成酵素 / 膜タンパク質 |
Research Abstract |
I developed the technology for incorporating non-natural amino acids into proteins in living cells, and applied it for protein researches. Thus, (i) target proteins were labeled in mammalian cells, (ii) a naturally-occurring posttranslational modification was incorporated into protein at defined sites, (iii) novel protein-protein interactions were discovered by in vivo photo-crosslinking method, and (iv) non-natural amino acids were site-specifically incorporated into proteins in insect cells for the first time.
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Report
(4 results)
Research Products
(42 results)
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[Journal Article] Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRNATyr pair by the archaeal tyrosine pair in Escherichia coli for genetic code expansion2010
Author(s)
Iraha, F., Oki, K., Kobayashi, T., Ohno. S., Yokogawa, T., Nishikawa, K., Yokoyama, S., Sakamoto, K.
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Journal Title
Nucleic Acids Res (in press)
Related Report
Peer Reviewed
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[Journal Article] Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRNA^<Tyr> pair by the archaeal tyrosine pair in Escherichia coil for genetic code expansion2010
Author(s)
Iraha, F., Oki, K., Kobayashi, T., Ohno, S., Yokogawa, T., Nishikawa, K., Yokoyama, S., Sakamoto, K.
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Journal Title
Nucleic Acids Research (in press)
Related Report
Peer Reviewed
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[Journal Article] Genetic encoding of 3-iodo-L-tyrosine in Escherichia coli for single-wavelength anomalous dispersion phasing in protein crystallography2009
Author(s)
Sakamoto, K., Murayama, K., Oki, K., Iraha, F., Kato-Murayama, M., Takahashi, M., Ohtake, K., Kobayashi, T., Kuramitsu, S., Shirouzu, M., Yokoyama S.
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Journal Title
Structure 17
Pages: 335-344
Related Report
Peer Reviewed
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[Journal Article] Genetic encoding of 3-iodo-L-tyrosine in Escherichia coli for single-wavelength anomalous dispersion phasing in protein crystallography2009
Author(s)
Sakamoto, K., Murayama, K., Oki, K., Iraha, F., Kato-Murayama, M., Takahashi, M., Ohtake, K., Kobayashi, T., Kuramitsu, S., Shirouzu, M., Yokoyama, S.
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Journal Title
Structure 17
Pages: 335-344
Related Report
Peer Reviewed
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[Journal Article] Multi-step engineering of pyrrolysyl-tRNA synthetase to genetically encode N-(o-azidobenzyl oxycarbonyl)lysine for site-specific protein modification2008
Author(s)
Yanagisawa, T., Ishii, R., Fukunaga, R., Kobayashi, T., Sakamoto, K., Yokoyama, S.
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Journal Title
Chem. Biol 15
Pages: 1187-1197
Related Report
Peer Reviewed
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[Journal Article] Adding L-lysine derivatives to the genetic code of mammalian cells with engineered pyrrolysyl-tRNA synthetases2008
Author(s)
Mukai, T., Kobayashi, T., Hino, N., Yanagisawa, T., Sakamoto, K., Yokoyama, S.
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Journal Title
Biochem. Biophys. Res. Commun 371
Pages: 818-822
Related Report
Peer Reviewed
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[Journal Article] Crystallographic studies on multiple conformational states of active-site loops in pyrrolysyl-tRNA synthetase2008
Author(s)
Yanagisawa, T., Ishii, R., Fukunaga, R., Kobayashi, T., Sakamoto, K., Yokoyama, S.
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Journal Title
J. Mol. Biol 378
Pages: 634-52
Related Report
Peer Reviewed
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[Journal Article] Multi-step engineering of pyrrolysyl-tRNA synthetase to genetically encode Nε-(o-azidobenzyloxycarbonyl)lysine for site-specific protein modification2008
Author(s)
Yanagisawa, T., Ishii, R., Fukunaga, R., Kobayashi, T., Sakamoto, K., Yokoyama, S.
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Journal Title
Chemistry & Biology 15
Pages: 1187-1197
Related Report
Peer Reviewed
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[Journal Article] Adding L-lysine derivatives to the genetic code of mammalian cells with engineered pyrrolysyl-tRNA synthetases2008
Author(s)
Mukai, T., Kobayashi, T., Hino, N., Yanagisawa, T., Sakamoto, K., Yokoyama, S.
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Journal Title
Biochemical and Biophysical Research Communications 371
Pages: 818-822
Related Report
Peer Reviewed
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[Journal Article] Crystallographic studies on multiple conformational states of active-site loops in pyrrolysyl-tRNA synthetase2008
Author(s)
Yanagisawa, T., Ishii, R., Fukunaga, R., Kobayashi, T., Sakamoto, K., Yokoyama, S.
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Journal Title
Journal of Molecular Biology 378
Pages: 634-52
Related Report
Peer Reviewed
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