High-resolution multi-dimensional NMR spectroscopy of proteins in human cells

Kohsuke Inomata, Ayako Ohno, Hidehito Tochio (), Shin Isogai, Takeshi Tenno, Ikuhiko Nakase, Toshihide Takeuchi, Shiroh Futaki, Yutaka Ito, Hidekazu Hiroaki and Masahiro Shirakawa ()
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Kohsuke Inomata: Graduate School of Engineering, Kyoto University, Nishikyo-Ku, Kyoto 615-8510, Japan
Ayako Ohno: Graduate School of Engineering, Kyoto University, Nishikyo-Ku, Kyoto 615-8510, Japan
Hidehito Tochio: Graduate School of Engineering, Kyoto University, Nishikyo-Ku, Kyoto 615-8510, Japan
Shin Isogai: Graduate School of Engineering, Kyoto University, Nishikyo-Ku, Kyoto 615-8510, Japan
Takeshi Tenno: CREST, and,
Ikuhiko Nakase: Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan
Toshihide Takeuchi: Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan
Shiroh Futaki: SORST, JST, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
Yutaka Ito: CREST, and,
Hidekazu Hiroaki: CREST, and,
Masahiro Shirakawa: Graduate School of Engineering, Kyoto University, Nishikyo-Ku, Kyoto 615-8510, Japan

Nature, 2009, vol. 458, issue 7234, 106-109

Abstract: In-cell NMR: protein structure in human cells Until now, in-cell NMR spectroscopy of living cells has been limited to bacteria and Xenopus laevis oocytes. Wider application to living eukaryote cells has been limited by the relatively inefficient delivery of isotope-labelled proteins into these cells. Now Inomata et al. show that it is possible to deliver suitably labelled proteins into the cytosol of human cells via the pyrenebutyrate-mediated action of cell-penetrating peptides that have been linked covalently to the proteins of interest. When the proteins are released by endogenous enzymes or autonomous reductive cleavage, high-resolution two-dimensional heteronuclear NMR spectra of proteins inside living human cells can be obtained. This technique could be a powerful tool for the design and screening of drugs targeted to intracellular proteins.

Date: 2009
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DOI: 10.1038/nature07839

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