| Project/Area Number |
16K07323
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Research Field |
Biophysics
|
|---|
| Research Institution | Kobe University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
木村 哲就 神戸大学, 理学研究科, 特命講師 (70506906)
武内 総子 神戸大学, 大学教育推進機構, 助教 (00448169)
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
|
|---|
| Keywords | 膜タンパク質 / 電子伝達 / ヘム / アポトーシス / ナノディスク / シトクロムb561 / アスコルビン酸 / 生体膜 / cytochrome b561 / モノデヒドロアスコルビン酸ラジカル / 癌抑制遺伝子 / タンパク質 |
|---|
| Outline of Final Research Achievements |
We have succeeded to perform large-scale expression and high purity purification of 101F6 protein using the 101F6 protein expression system by imploying the methanol-utilizing yeast Pichia pastoris. Cell extracts from human cancer tissue-derived A549 cells were prepared using surfactant DDM or β-OG. After thoroughly mixing the cell extract with the purified 101F6 protein, followed by by magnetic beads antibody method using antibodies that specifically recognize and bind the 101F6 protein, we obtained a group of proteins that seems to interact with the 101F6 protein in human cells by SDS-PAGE and silver staining analyses. Then we have succeeded in reconstituting the purified 101F6 protein into nanodiscs which are very good models of biological membranes, Further, we have succeeded in measuring the enzyme activity of purified 101F6 protein for the first time.
|
| Academic Significance and Societal Importance of the Research Achievements |
本研究により、101F6タンパク質はferric reductase活性を持つ事が既に報告されているDcytbと同様にferric reductase活性を持つ事が初めて証明された。この結果は、101F6の持つcaspase非依存性アポトーシス誘導作用がferric reductase 活性と何らかの関連を持つ事を示すものと考えられる。また、本研究において初めて精製したb561タンパク質ホモログそのものを用いてferric reductase酵素活性を直接に測定できる事を示した。そういう意味で本研究の意義は非常に大きい
|
