| Project/Area Number |
20K22626
|
|---|
| Research Category |
Grant-in-Aid for Research Activity Start-up
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
0701:Biology at molecular to cellular levels, and related fields
|
|---|
| Research Institution | Kanazawa University |
|---|
Principal Investigator |
Marchesi Arin 金沢大学, ナノ生命科学研究所, 特任助教 (80882240)
|
|---|
| Project Period (FY) |
2020-09-11 – 2022-03-31
|
| Project Status |
Discontinued (Fiscal Year 2021)
|
| Budget Amount *help |
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
|
|---|
| Keywords | ion channels / gating / atomic force microscopy / membrane proteins / CNG channels / Atomic Force Microscopy / Ion channels / Purification / Gating |
|---|
| Outline of Research at the Start |
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.
|
| Outline of Annual Research Achievements |
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.
|
