|Budget Amount *help
¥153,010,000 (Direct Cost : ¥117,700,000、Indirect Cost : ¥35,310,000)
Fiscal Year 2013 : ¥25,350,000 (Direct Cost : ¥19,500,000、Indirect Cost : ¥5,850,000)
Fiscal Year 2012 : ¥25,350,000 (Direct Cost : ¥19,500,000、Indirect Cost : ¥5,850,000)
Fiscal Year 2011 : ¥20,280,000 (Direct Cost : ¥15,600,000、Indirect Cost : ¥4,680,000)
Fiscal Year 2010 : ¥70,980,000 (Direct Cost : ¥54,600,000、Indirect Cost : ¥16,380,000)
Fiscal Year 2009 : ¥11,050,000 (Direct Cost : ¥8,500,000、Indirect Cost : ¥2,550,000)
Equilibrium shifting from the unbound states to the bound states is useful to obtain structural and dynamic information of transient protein complexes at atomic resolutions, in particular, the complexes with large amplitude motions inside. We developed two techniques, molecular tethering (via inter-molecular disulfide bonds) and molecular stiffening (via intra-molecular disulfide bonds), for this purpose. We also proposed intentional creation of space in protein crystal lattice, and place a target region in the space created. The key technique is the rigid connection to fix the relative orientation of the protein of interest and a tag protein. These new techniques were applied to analyze the dynamics of a mitochondrial presequence peptide as bound to the mitochondrial receptor Tom20, and the stabilization of oligosaccharyltransferase-peptide complexes for crystallization.