Constructing Nanoscale Molecule Sorters Powered by F1-ATPase Motors

构建由 F1-ATPase 电机驱动的纳米级分子分选机

• 批准号: 0084732
• 负责人: Carlo Montemagno
• 金额: --
• 依托单位: Cornell Univ - State: AWDS MADE PRIOR MAY 2010
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0084732&HistoricalAwards=false
• 关键词: Constructing; Nanoscale; Molecule; Sorters; Powered

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

本研究的主要目标是构建一个由 F1-ATPase 驱动的纳米级分子分选器（见下图）。作为该总体目标的一部分，已确立了五个里程碑目标：1.) 评估影响目标分子与收集器结合/洗脱的物理和化学特性；2.) 设计和制造分子分选器的收集器部分，并证明其特异性结合分子的能力；3.) 设计和制造收集器的存储部分；4.) 将 F1-ATP 酶与制造的结构集成，以 组装最终的分子分选机； 5.) 演示集成分子分选器的功能，并评估其性能和效率。分子分选器将由两部分组成：主动收集器和连接的存储单元。工程纳米级收集器将涂有a-绿色荧光蛋白（GFP）IgG 分子，并将由位于镍柱中的F1-ATPase 电机驱动。该收集器单元将从周围溶液中去除 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。内衬电极的连接导管将产生电场梯度，从而充当渗透泵以将 GFP 浓缩在末端储存器中。将使用末端储存器的荧光成像来评估收集的速率和效率。该装置的构建将建立一个适应性强的系统，用于构建基于细胞内的分子分选器，能够直接连接和利用细胞生理反应