| 研究課題/領域番号 |
20K22626
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| 研究種目 |
研究活動スタート支援
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| 配分区分 | 基金 |
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| 審査区分 |
0701:分子レベルから細胞レベルの生物学およびその関連分野
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| 研究機関 | 金沢大学 |
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研究代表者 |
Marchesi Arin 金沢大学, ナノ生命科学研究所, 特任助教 (80882240)
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| 研究期間 (年度) |
2020-09-11 – 2022-03-31
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| 研究課題ステータス |
中途終了 (2021年度)
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| 配分額 *注記 |
2,860千円 (直接経費: 2,200千円、間接経費: 660千円)
2021年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
2020年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
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| キーワード | ion channels / gating / atomic force microscopy / membrane proteins / CNG channels / Atomic Force Microscopy / Ion channels / Purification / Gating |
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| 研究開始時の研究の概要 |
The proposal will advance our understanding of ion channels function by exploiting a recently developed mechanical microscope in combination to advanced optics. The combination of these techniques will allow us to study ion channels structural dynamcs and link it to their physiological function.
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| 研究実績の概要 |
The SthK channel gene (UniProtKB G0GA88)was subcloned in the appropriate vector (pCGFP-BC) for overexpression in bacteria (BL21-DE3). Channel expression, purification and reconstitution in artificial bilayer was implemented successfully. The sample was characterized by means of force-spectroscopy (i.e. adsorbed to the AFM probe and unfolded from the membrane bilayer) and high-speed AFM imaging both in the presence of the endogenous ligand (cAMP) and its absence. This allowed us to define key interactions among the different protein domains and the overall mechanisms of gating (i.e. the opening of the permeation pathway). One paper, in collaboration with the ISAS-Italy (International School for Advanced Studies) is currently in revision and another is in preparation.
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