Protein structure determination in living cells by in-cell NMR spectroscopy

Daisuke Sakakibara, Atsuko Sasaki, Teppei Ikeya, Junpei Hamatsu, Tomomi Hanashima, Masaki Mishima, Masatoshi Yoshimasu, Nobuhiro Hayashi, Tsutomu Mikawa, Markus Wälchli, Brian O. Smith, Masahiro Shirakawa, Peter Güntert and Yutaka Ito ()
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Daisuke Sakakibara: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Atsuko Sasaki: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Teppei Ikeya: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Junpei Hamatsu: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Tomomi Hanashima: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Masaki Mishima: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Masatoshi Yoshimasu: Cellular and Molecular Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan
Nobuhiro Hayashi: Institute for Comprehensive Medical Science, Fujita Health University, Toyoake-shi, Aichi 470-1192, Japan
Tsutomu Mikawa: Research Group for Bio-supramolecular Structure-Function, RIKEN, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan
Markus Wälchli: Bruker BioSpin, 3-21-5 Ninomiya, Tsukuba-shi, Ibaraki 305-0051, Japan
Brian O. Smith: Faculty of Biomedical and Life Sciences, University of Glasgow
Masahiro Shirakawa: CREST/Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
Peter Güntert: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan
Yutaka Ito: Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji, Tokyo 192-0397, Japan

Nature, 2009, vol. 458, issue 7234, 102-105

Abstract: In-cell NMR: three-dimensional protein structures Previously, the relatively low sensitivity of spectroscopy and short lifetime of the samples have made it difficult to acquire sufficient structural information to determine protein structures using in-cell NMR spectroscopy. It can take one or two days to collect NMR data, too long for living cells. Sakakibara et al. overcome this limitation by collecting enough data in a few hours; they report the first three-dimensional protein structure determined exclusively on the basis of information obtained in living Escherichia coli. The model protein used in this 'proof of principle' study is the putative heavy metal-binding protein TTHA1718 from Thermus thermophilus.

Date: 2009
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DOI: 10.1038/nature07814

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