EAGER/Collaborative Research: Large Scale Microtubule-Based Nanomanufacturing of Single Kinesin Patterns with Ultrahigh Resolution

该探索性研究早期概念资助 (EAGER) 项目的目标是开发一种以超高分辨率（低于 10 nm）对单一蛋白质进行图案化的新概念。该方法基于自组装和自我识别，起源于过于拥挤的细胞内环境，不允许扩散成为细胞质内物质运动的有效机制。具体来说，垂直固定在原子力显微镜尖端的原位聚合微管（细胞骨架丝）用作纯化驱动蛋白分子马达的轨道；在三磷酸腺苷水解产生的化学能作用下，驱动蛋白从微管尖端沉积到​​紧邻的玻璃表面上，形成单一蛋白质的纳米阵列。 EAGER 开发的这项技术在药物输送、筛选、纳米电子学和纳米传感器等领域造福社会。除了通过生物分子可用于超高分辨率打印纳米阵列的原理证明认识到这项技术的价值之外，这项研究还提供了对单个纳米材料（有机和无机）进行图案化的解决方案。固有的跨学科性质为吸引和整合具有跨学科教育经验的学生提供了巨大的机会（该项目将雇用两名毕业生）。基于仿生的纳米制造领域的进步将被纳入西弗吉尼亚大学的细胞机器和 RPI 的生物材料加工两门课程中。最后，PI 将利用西弗吉尼亚大学生物工程师学会制作廉价的海报来宣传生物纳米技术的进步，从而为纳米技术的公共教育和对代表性不足的人群（即妇女、农村社区）的宣传做出贡献。

The objective of this Early-Concept Grant for Exploratory Research (EAGER) project is to develop a novel concept for patterning single protein with ultrahigh resolution (below 10 nm). The approach is based on self-assembly and self-recognition and originates in the intracellular environment that is too crowded to allow diffusion to be an efficient mechanism for the movement of materials within the cytoplasm. Specifically, in situ polymerized microtubule (cytoskeletal filaments) affixed vertically to the tip of an atomic force microscope serves as track for purified kinesin molecular motors; under the chemical energy derived from adenosine triphosphate hydrolysis kinesin is deposited from the microtubule tip onto a glass surface situated in close proximity leading to nanoarrays of single protein. This EAGER-developed technology benefits society in areas as: drug delivery, screening, nanoelectronics, and nanosensors. Beyond recognizing the value of this technology through the proof of principle that biological molecules can be used for printing nanoarrays with ultrahigh resolution, this research also provides solutions to patterning individual nanomaterial (both organic and inorganic). The inherent interdisciplinary nature offers tremendous opportunities for enticing and integrating students with educational experience across diverse disciplines (two graduates will be employed by this program). The advances in the field of biomimetic-based nanomanufacturing will be incorporated in two courses Cellular machines at WVU and Processing of Biomaterials at RPI. Lastly, the PI will use Society for Biological Engineers at WVU to popularize bionanotechnology by generating inexpensive posters highlighting the advances and thus contributing to public education in nanotechnology and outreach to underrepresented populations, (i.e., women, rural communities).