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

这个探索性研究早期概念资助（EAGER）项目的目标是开发一种新的概念，以10 nm以下的分辨率对单个蛋白质进行图案化。该方法基于自组装和自识别，并且起源于细胞内环境，该细胞内环境太拥挤而不允许扩散成为细胞质内物质移动的有效机制。具体地说，原位聚合的微管（细胞骨架丝）垂直固定在原子力显微镜的尖端，作为纯化的驱动蛋白分子马达的轨道;在来自三磷酸腺苷水解的化学能作用下，驱动蛋白从微管尖端沉积到位于附近的玻璃表面上，从而形成单一蛋白质的纳米阵列。EAGER开发的这项技术在以下领域造福社会：药物输送，筛选，纳米电子学和纳米传感器。除了通过证明生物分子可用于打印分辨率为1000的纳米阵列的原理来认识到这项技术的价值外，这项研究还提供了图案化单个纳米材料（有机和无机）的解决方案。固有的跨学科性质为吸引和整合具有不同学科教育经验的学生提供了巨大的机会（该计划将雇用两名毕业生）。仿生纳米制造领域的进展将被纳入两门课程WVU的细胞机器和RPI的生物材料加工。最后，PI将利用西弗吉尼亚大学的生物工程师协会，通过制作廉价的海报来推广生物纳米技术，突出进步，从而促进纳米技术的公共教育和对代表性不足的人群的宣传，（即，农村社区）。

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).