CAREER: Elucidation of Surface Affects on Biochemical Reactions

华盛顿大学的Daniel Chiu教授因研究受限空间对酶动力学的影响而获得了分析和表面化学项目的CAREER奖。单分子光谱将用于检测酶，如β -半乳糖苷酶作用于100纳米合成囊泡和纳米通道中设计的平面支撑双层中含有的荧光团。电穿孔和微流体将用于操作囊泡和单酶。这些研究将确定与细胞纳米环境相关的表面电荷和疏水性是否会对蛋白质的活性产生重大影响。这些研究将有助于更好地理解细胞功能和区隔化。PI将提供一门新的研究生课程，涵盖微加工、流体力学、傅立叶光学和生物分析方法等主题，并将广泛使用基于网络的技术来辅助普通化学课程。大多数蛋白质反应研究都是在烧杯中的溶液中进行的，细胞表面的任何影响都没有了。单分子光谱学的出现使得研究微小通道或囊泡内的单个酶分子成为可能，模仿细胞中封闭的超小空间。在之前的研究中，分子与分子之间的差异已经显现出来，这些差异可能是显著的。然后可以研究壁和有限空间对生化反应的影响。从事本研究和研究生课程中使用的技术的学生可以将知识转移到许多工作中，如芯片实验室和传感器。

Professor Daniel Chiu of the University of Washington has received a CAREER award from the Analytical and Surface Chemistry Program to investigate the effects of constrained space on enzyme kinetics. Single molecule spectroscopy will be utilized to examine enzymes such as beta-galactosidase acting on fluorophores contained in 100 nm synthetic vesicles and designed planar supported bilayers in nanochannels. Electroporation and microfluidics will be used for manipulation of vesicles and single enzymes. The studies will ascertain whether the surface charge and hydrophobicity associated with cellular nanoenvironments can exert significant influence on the activity of proteins. These studies will lead to a better understanding of cellular functioning and compartmentalization. The PI will offer a new graduate course covering topics in microfabrication, fluid mechanics, Fourier optics and bioanalytical methods, and also will use web-based technologies extensively to assist in the General Chemistry courses. Most protein reaction studies are carried out in solutions in beakers, and any effects of the cell surfaces are lost. The advent of single molecule spectroscopy makes possible the study of single enzyme molecules inside tiny channels or vesicles, mimicking the enclosed ultrasmall spaces found in cells. Differences from molecule to molecule have shown up in previous studies, and these can be significant. The effects of the walls and limited spaces on the biochemical reactions can then be studied. Students engaged in the technologies used in this research and in the graduate course can transfer the knowledge to many efforts such as lab-on-a-chip and sensors.