A Gel-Encapsulated Bioreactor System for NMR Studies of Protein–Protein Interactions in Living Mammalian Cells

Authors

  • Satoshi Kubo,

    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author
  • Dr. Noritaka Nishida,

    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author
  • Yuko Udagawa,

    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author
  • Osamu Takarada,

    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author
  • Dr. Shinji Ogino,

    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author
  • Prof. Dr. Ichio Shimada

    Corresponding author
    1. Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    2. Biomedicinal Information Research Center(BIRC), National Institute of Advanced Industrial Science and Technology (AIST), 2-42 Aomi, Koto-ku, Tokyo 135-0064 (Japan)
    • Division of Physical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
    Search for more papers by this author

  • This work was supported by a grant from the Japan New Energy and Industrial Technology Development Organization (NEDO) and the Ministry of Economy, Trade, and Industry (METI; to I.S.). S.K. is a research fellow of the Japan Society for the Promotion of Science (JSPS).

Abstract

original image

Staying alive: The major limitation of in-cell NMR spectroscopy methods is the occurrence of cell death during the NMR measurement. To overcome this problem, a bioreactor was utilized that can perfuse the cells in the NMR tube, thereby maintaining the conditions for more than 5 h. By using the bioreactor, the binding site of an externally introduced protein for an endogenous molecule in HeLa S3 cells was identified.

Ancillary