2021 Fiscal Year Annual Research Report
Nanoscale probing and visualization of cyclic nucleotide-gated ion channels structural dynamics and function
| Project/Area Number |
20K22626
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| Research Institution | Kanazawa University |
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Principal Investigator |
Marchesi Arin 金沢大学, ナノ生命科学研究所, 特任助教 (80882240)
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| Project Period (FY) |
2020-09-11 – 2022-03-31
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| Keywords | ion channels / gating / atomic force microscopy / membrane proteins / CNG channels |
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| 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.
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Research Products
(3 results)
