Structural and functional analyses of autophagy essencial protein Atg9

Research Project

Project/Area Number	15K21608
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Structural biochemistry Cell biology
Research Institution	Microbial Chemistry Research Foundation
Principal Investigator	MATOBA Kazuaki 公益財団法人微生物化学研究会, 微生物化学研究所・構造生物学研究部, 研究員 (60613792)
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000) Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000) Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords	オートファジー / 膜タンパク質
Outline of Final Research Achievements	To determine a structure of autophagy essential membrane protein, overexpression and solubilization conditions were established and the purity was high enough to be detected as a single band using CBB staining. Protein-Fab complex crystallization was performed, but structural analysis has not been the success. In order to observe the single particle by cryo-electron microscope, Nanodisc-protein-Fab complex was reconstituted. As a result, Fab bound particle was observed by negative staining.