| 研究概要 |
Development of the proposed project following the original research plan proved to be extremely challenging, especially research experiments related to the coupling of synthesized Fe_3O_4 nanoparticles with the biological material (the kinesin protein).
Initial experiments were carried out to synthesize Fe_3O_4 nanoparticles bearing a free carboxyl surface. Even though nanoparticles were successfully prepared, coupling to the kinesin protein could not be achieved, as the environmental medium required for the coupling reaction would lead to nanoparticle degradation. As such environmental medium is compatible with amine-terminated nanomaterial, Fe_3O_4 nanoparticles were synthesized to counter this problem. Unfortunately, the synthesis could not be achieved, as nanop articles conglomeration would take place. Several different strategies were tried in order to synthesized magnetic iron oxide nanoparticles bearing a primary amine surface, and such research is still being conducted.
The latest approach being currently studied to couple Fe_3O_4 nanoparticles with kinesin protein is the bio-functionalization of the magnetic nanomaterial. In this method, nanoparticles are encapsulated in biotin-tagged micelles and coupled to the biotinylated kinesin protein via an avidin liker.
Such difficulties, which were essential to the proposed project, and more importantly, an prerequisite to further development, yielded a significant delay in the project.
|
| 現在までの達成度 (区分) |
現在までの達成度 (区分)
4: 遅れている
理由
The kinesin protein with a terminal biotin tag expressed in E. coli cells proved to be functional. However, magnetic material could not be loaded to the protein in order to induce electric impulses via its circular motion (even though, commercially available quantum dots could be successfully attached to the protein and its motion over microtubule tracks were observed under fluorescence microscopy). Different approaches are currently being studied to solve this problem.
|
| 今後の研究の推進方策 |
In order to achieve a proper advancement of the proposed project, the crucial iron oxide nanoparticle synthesis must be solved. It is unclear which is the best approach to couple synthetic nanomaterial to biological material. Medium compatibility proved to be a challenge, and although the current bio-functionalization method using micelles is proving to be promising, it is very time consuming. Therefore, having a amine-terminated nanoparticle surface would be an elegant alternative to save time and couple nanoparticles to protein in fewer steps. Unfortunately, we were not able at the present time to achieve such task.
|
