CAREER: The Role of Ribosome Conformational Dynamics on the Mechanism of Protein Synthesis

这个项目关注的是最基本的细胞过程之一：核糖体的蛋白质合成，核糖体是一种普遍保守的核糖核蛋白酶。冈萨雷斯实验室的最终目标是表征参与蛋白质合成的核糖体和其他生物分子的构象动力学，并利用这些数据来阐明蛋白质合成的详细分子机制。新的荧光光谱方法被用来在单分子水平上研究核糖体分子机，重点是生物分子的构象动态区域，其功能重要性在很大程度上仍然是个谜。利用单分子荧光，核糖体内构象变化的机制被识别、量化并与蛋白质合成的整个过程中的生化步骤相耦合。然而，要达到这种水平的分辨率，需要合并分子生物学、生物化学、生物物理学、光学、显微镜、表面化学、微流体和统计数据分析等领域。因此，该实验室是高度跨学科的，并积极参与补充技术开发，例如将微流控设计原则应用于用于单分子荧光测量的样品池，并开发用于数据分析的数学上可靠的统计模型，这将扩大单分子荧光光谱仪的能力。除了在蛋白质合成领域的独家影响，这项研究开发的一般原理直接适用于对细胞具有关键功能重要性的众多复杂生化反应的研究。冈萨雷斯实验室研究的跨学科性质为整合不同科学背景的学生和促进跨学科培训提供了独特的机会。为了在哥伦比亚大学更广泛的院系和大学社区中产生类似的教育影响，冈萨雷斯教授正在将核酸和单分子生物物理学整合到课程中，参与设计一个共享的分子生物学设施，并为该设施的用户组织一个论坛。冈萨雷斯教授还制定了一项长期的本科生/高中生研究计划，由本科生和高中生研究人员专门执行的项目组成。这种跨学科研究项目各方面的教育和实践经验将使学生为未来在当代科学实验室的工作做好准备。该项目由分子、细胞生物学和化学分部共同资助。

This project focuses on one of the most fundamental of cellular processes: protein synthesis by the ribosome, a universally conserved ribonucleoprotein enzyme. The ultimate goal in the Gonzalez laboratory is to characterize the conformational dynamics of the ribosome and other biomolecules involved in protein synthesis and utilize these data to elucidate the detailed molecular mechanism of protein synthesis. Novel fluorescence spectroscopic approaches are used to study the ribosomal molecular machine at the single-molecule level, focusing on conformationally dynamic regions of the biomolecule whose functional importance remains largely mysterious. Using single-molecule fluorescence, the mechanics of conformational changes within the ribosome are identified, quantified and coupled to biochemical steps in the overall process of protein synthesis. Achieving this level of resolution, however, requires merging the areas of molecular biology, biochemistry, biophysics, optics, microscopy, surface chemistry, microfluidics, and statistical data analysis. The laboratory is therefore highly interdisciplinary and is actively engaged in complementary technological developments, such as the application of microfluidic design principles to the sample cells used for single-molecule fluorescence measurements and the development of mathematically robust statistical models for data analysis that will expand the capabilities of single-molecule fluorescence spectroscopies. Beyond exclusive impact in the area of protein synthesis, the general principles developed by this research are directly applicable to the study of the numerous complex biochemical reactions that are of critical functional importance to the cell. The interdisciplinary nature of the research in the Gonzalez laboratory provides a unique opportunity to integrate students of varied scientific backgrounds and stimulate cross-discipline training. In order to achieve similar educational impact in the broader departmental and university communities at Columbia University, Prof. Gonzalez is integrating nucleic acid and single-molecule biophysics into the curriculum, participating in the design of a shared molecular biology facility, and organizing a discussion forum for users of this facility. Prof. Gonzalez has also put in place a long-term undergraduate/high-school student research program composed of projects to be exclusively executed by undergraduate and high-school student researchers. This educational and practical experience in all aspects of an interdisciplinary research project will prepare students for future work in contemporary scientific laboratory settings. This project is cofunded by the Divisions of Molecular and Cellular Biology and Chemistry.