SEMINAR

Crystallography using Synchrotron Radiation

Abstract
Biological macromolecules, such as proteins, and their assemblies play significant roles in many biological reaction systems. Detailed understanding of the functions of these molecules requires information derived from three-dimensional atomic structures. X-ray crystallography is one of the most powerful techniques to determine the three-dimensional structures of molecules at atomic level. However, it is usually very difficult to obtain good diffraction data from these crystals. Institute for Protein Research (IPR) of Osaka University is running a synchrotron radiation beamline for crystal structure analysis of biological macromolecular assemblies at SPring-8 (BL44XU) [1]. This beamline is designed to collect high quality diffraction data from biological macromolecule assembly crystals with large unit cells.

I will present the IPR beamline for biological macromolecular assemblies and its application to the structure determination of the voltage-dependent proton channel [2].

References

[1] A. Higashiura et al. (2015) SPring-8 BL44XU, beamline designed for structure analysis of large biological macromolecular assemblies. J. Synchrotron Rad., under review.
[2] K. Takeshita, et al. (2014) X-ray crystal structure of voltage-gated proton channel. Nat. Struct. Mol. Biol., 21, 352-357.

BIOGRAPHY

2003-present: Professor, Institute for Protein Research, Osaka University, Japan
1999-2003: Associate Professor, Institute for Protein Research, Osaka University, Japan
1995-1999: Associate Professor, Graduate School of Science, Hokkaido University, Japan
1994-1995: Visiting Scientist, Laboratory of Molecular Biology, Medical Research Council, UK
1986-1995: Assistant Professor, Photon Factory, National Laboratory for High Energy Physics, Japan
1999: PhD, Graduate School of Science, Osaka University, Japan

Research Fields and Interests:
Structure and structural changes of proteins in solution underpin function and play a critical role in drug development. We are developing methods for structural biology in solution, in particular by NMR and EPR spectroscopy. This includes the use of cell-free protein synthesis for selective isotope labelling and the incorporation of genetically encoded unnatural amino acids as specific sites for chemical tags, for selective detection by NMR spectroscopy, generation of pseudocontact shifts and chemical cross-linking.

Selected Publications:
Takeshita, K., Sakata, S., Yamashita, E., Fujiwara, Y., Kawanabe, A., Kurokawa, T., Okochi, Y., Matsuda, M., Narita, H., Okamura, Y., Nakagawa, A. (2014) X-ray crystal structure of voltage-gated proton channel. Nat. Struct. Mol. Biol., 21, 352-357.

Sasanuma, H., Tawaramoto, M. S., Lao, J. P., Hosaka, H., Sanda, E., Suzuki, M., Yamashita, E., Hunter, N., Shinohara, M., Nakagawa, A., Shinohara A. (2013) A new protein complex promoting the assembly of Rad51 filaments. Nat. Commun., 4, 1676.

Takeshita, K., Suetake, I., Yamashita, E., Suga, M., Narita, H., Nakagawa, A., Tajima, S. Structural insight into maintenance methylation by mouse DNA methyltransferase 1 (Dnmt1). Proc. Natl. Acad. Sci. USA, 108, 9055-9059.

Matsuda, M., Takeshita, K., Kurokawa, T., Sakata, S., Suzuki, M., Yamashita, E., Okamura Y., Nakagawa, A. (2011) Crystal structure of the cytoplasmic phosphatase and tensin homolog (PTEN)-like Region of Ciona intestinalis Voltage-sensing Phosphatase Provides Insight into Substrate Specificity and Redox Regulation. J. Biol. Chem., 286, 23368-23377 (2011).

Higashiura, A., Kurakane, T., Matsuda, M., Suzuki, M., Inaka, K., Sato, M., Kobayashi, T., Tanaka, T., Tanaka, H., Fujiwara K., Nakagawa, A. (2010) High-resolution X-ray crystal structure of bovine H-protein at 0.88 Å resolution. Acta Cryst. sect. D, A., 698-708.