Electron microscopy of structure and localization of signal processing complexes

2014 Fiscal Year Final Research Report

• Project Area: Structural basis of cell-signalling complexes mediating signal perception, transduction and responses
• Project/Area Number: 22121004
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: SATO Chikara 独立行政法人産業技術総合研究所, バイオメディカル研究部門, 研究グループ長 (00357146)
• Co-Investigator(Renkei-kenkyūsha): KAWATA Masaaki 独立行政法人産業技術総合研究所, 人工知能研究センター, 主任研究員 (20356843)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Keywords: シグナル伝達 / 分子複合体 / 電子顕微鏡 / タンパク質構造 / イオンチャンネル

Outline of Final Research Achievements

Protein complexes are essential for various kinds of physiological functions in the human body. By developing single particle analysis (SPA) using Transmission electron microscopy (TEM), the three-dimensional (3D) structure of signal processing protein complexes. Further, these structures including SecDF are interpreted by docking partial structure determined by X-ray crystallography.
The new atmospheric scanning electron microscope (ASEM) observes samples through electron-transparent film. Therefore, samples can be set in solution under an open atmosphere. Using this system, immuno-ASEM was successfully developed, and we have observed intracellular super-molecular-complex formations, including accumulation of STIM-1 in response to Ca2+ store depletion. Moreover, the observation method of small protein crystals are developed, and contributed to X-ray crystallography. Further, by multi-windowning ASEM, the observation efficiency was improved.

Free Research Field

構造細胞生物学