B01-2：In-cell NMR studies on metalloproteins for understanding biometal behaviours
Yutaka Ito
（Department of Chemistry, Tokyo Metropolitan University）
Lab website
Biomacromolecules occupy a significant fraction of the intracellular volume, resulting in molecular crowding. Metalloproteins at work in cells exhibit their biological functions through the excluded-volume effect, specific and non-specific interactions, various dynamical intracellular processes, and the binding to various biometal ions. In this research area we are going to perform in situ analysis of various intracellular events related to metalloproteins by employing “in-cell NMR spectroscopy”, which is currently the only tool to observe bio-macromolecules at atomic resolution in living biological systems. Specific research themes are as follows. First, we will establish a versatile and robust in-cell NMR platform applicable to various studies on metalloproteins. Furthermore, we will contribute to this research area by providing other researchers with state-of-the-art solution NMR methods, including advanced procedures for NMR measurement and analysis, protein 3D structure determination using Bayesian inference, etc.
Major publications
T. Tanaka, *T. Ikeya, H. Kamoshida, Y. Suemoto, M. Mishima, M. Shirakawa, P. Güntert, and *Y. Ito
“High-Resolution Protein 3D Structure Determination in Living Eukaryotic Cells”
Angewandte Chemie International Edition, 2019, 58, 7284-7288
doi: 10.1002/anie.201900840
*T. Ikeya, T. Hanashima, S. Hosoya, M. Shimazaki, S. Ikeda, M. Mishima, P. Güntert, and *Y. Ito
“Improved in-cell structure determination of proteins at near-physiological concentration”
Scientific Reports, 2016, 6, 38312
doi: 10.1038/srep38312
Y. Hikone, G. Hirai, M. Mishima, K. Inomata, T. Ikeya, S. Arai, M. Shirakawa, M. Sodeoka, and *Y. Ito
“A new carbamidemethyl-linked lanthanoid chelating tag for PCS NMR spectroscopy of proteins in living HeLa cells”
Journal of Biomolecular NMR, 2016, 66, 99-110
doi: 10.1007/s10858-016-0059-4
J. Hamatsu, D. O’Donovan, T. Tanaka, T. Shirai, Y. Hourai, T. Mikawa, T. Ikeya, M. Mishima, W. Boucher, B.O. Smith, E.D. Laue, M. Shirakawa, and *Y. Ito
“High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System”
Journal of American Chemical Society, 2013, 135, 1688-1691
doi: 10.1021/ja310928u
D. Sakakibara, A. Sasaki, T. Ikeya, J. Hamatsu, T. Hanashima, M. Mishima, M. Yoshimasu, N. Hayashi, T. Mikawa, M. Wälchli, B.O. Smith, M. Shirakawa, P. Güntert, and *Y. Ito
“Protein structure determination in living cells by in-cell NMR spectroscopy”
Nature, 2009, 458, 102–105
doi: 10.1038/nature07814