Constructing Nanoscale Molecule Sorters Powered by F1-ATPase Motors

这项研究的主要目标是构建一个由F1-ATPase驱动的纳米级分子分离器（见下图）。作为这一总体目标的一部分，已经建立了五个里程碑目标：1.评估影响目标分子与收集器结合/洗脱的物理和化学性质;2.）设计并制作了分子分选仪的收集器部分，并证明其特异性结合分子的能力;3.）设计并制作收集器的存储部分;4.）将F1-ATP酶与制造的结构整合以组装最终的分子分离器;以及5.）演示集成分子分选仪的功能，并评估其性能和效率。分子分选仪将由两部分组成：活性收集器和连接的存储单元。这种纳米级的收集器将涂上一层绿色荧光蛋白（GFP）IgG分子，并由镍柱上的F1-ATP酶发动机驱动。这个收集器单元将从周围的溶液中去除GFP，并通过液体界面处的疏水性和电势的变化将它们转移到存储单元（包含油基介质）中。连接导管内排列着电极，会产生一个电梯度，从而起到渗透泵的作用，将GFP集中在末端的储库中。收集的速率和效率将使用末端水库的荧光成像进行评估。该装置的构建将建立一个可直接连接和利用细胞生理反应的细胞内分子分选系统

The primary objective of the proposed research is to construct a nanoscale moleculesorter powered by F1-ATPase (see illustration below). Five milestone goals have beenestablished as part of this overall objective:1.) Evaluate the physical and chemical properties affecting the binding/elution oftarget molecules to the collector;2.) Design and fabricate the collector portion of the molecule sorter, and demonstrateits ability to specifically bind molecules;3.) Design and fabricate the storage portion of the collector;4.) Integrate of F1-ATPase with fabricated structures to assemble the final moleculesorter; and5.) Demonstrate the functionality of the integrated molecule sorter, and evaluate itsperformance and efficiency.The molecule sorter will consist of two parts: the active collector and the connectedstorage units. The engineered nanoscale collector will be coated with a-green fluorescentprotein (GFP) IgG molecules, and will be actuated by F1-ATPase motors positioned innickel posts. This collector unit will remove GFP from the surrounding solution, andtransfer them into the storage units (containing an oil-based medium) through changes inhydrophobicity and electrical potential at the liquid interface. Connective conduits linedwith electrodes will generate an electric gradient, and thus act as an osmotic pump toconcentrate GFP in the end reservoirs. The rate and efficiency of collection will beevaluated using fluorescent imaging of the end reservoirs. Construction of this devicewill establish an adaptable system for constructing intracellular-based molecule sorterscapable of directly interfacing and utilizing cell physiological responses