| Budget Amount *help |
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2014: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2012: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000)
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| Outline of Final Research Achievements |
Human membrane proteins (hMPs) are important targets in the structural biology field because they contain a lot of drug targets. However, expression of recombinant hMPs by conventional prokaryotic cells is usually difficult. In this study, we constructed a robust strategy to express hMPs using two yeast species, P. pastoris and K. lactis. We further developed a variety of methods to isotopically label recombinant hMPs for structural studies using nuclear magnetic resonance (NMR) spectroscopy. We expressed six hMPs in the yeast membranes, and some of them were labeled with 2H, 15N, and 13C in a methyl-group selective manner. Using these samples, we could obtain molecular properties and structural information of several hMPs. The protocol established in this study is a widely applicable method and will contribute to following NMR studies of hMPs.
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