FIBR: How Do Proteins Fold Into Their Native and Functional Structures In-Vitro and in The Physiological Milue of The Living cell?

在所有活细胞中，蛋白质自组装或折叠成具有独特功能的精确三维结构，这些结构来自由核糖体（负责蛋白质合成的酶）组装的线性多肽链。 蛋白质的精确折叠是由其DNA序列决定的，但研究人员尚未破译序列编码结构的规则（“蛋白质折叠问题”）。 解决这个问题对于理解基因序列变异如何转化为蛋白质和细胞功能的变异至关重要。 控制和引导折叠过程的结构动力学的力的微妙平衡对细胞内的环境条件高度敏感。 该项目的目标是协同应用尖端方法，包括单分子光谱学，超快微流体混合，光诱导电子转移，非天然氨基酸标记，线粒体蛋白质运输，化学肽合成和模拟建模使用分布式和超级计算系统，在模拟活细胞内自然折叠环境的条件下研究蛋白质折叠。研究人员将在各种条件下研究三种不同蛋白质在简单溶液（体外）中的未折叠状态，同时蛋白质直接在核糖体本身上制备。通过将这些研究与在线粒体基质的拥挤环境（细胞内折叠环境的模拟物）中进行的蛋白质折叠实验进行比较，该项目旨在了解体外和体内折叠环境之间的主要差异以及这些差异对蛋白质折叠机制的影响。 该项目将通过开发新的工具和方法以及在分子水平上研究复杂生物过程的一般方法对细胞生物学领域产生广泛影响。 一个推广方案将把这些研究工具传播给代表性不足的机构的教师和学生，并加强中等教育一级的科学和技术知识。 该项目代表了由Shimon韦斯领导的加州大学洛杉矶分校研究人员的跨学科合作，子奖项授予斯坦福大学（Vijay Pande）、德克萨斯农工大学（亚瑟约翰逊）、加州大学戴维斯分校（Olgica Bakajin）、密歇根州立大学（丽莎拉皮德斯）和斯克里普斯研究所（Jeff Kelly）。 大量的学生和博士后研究员将在化学，生物物理学和模拟的界面上接受研究的概念和技术方面的高级培训。

In all living cells, proteins self-assemble or fold into precise, three dimensional structures having unique functions from a linear polypeptide chain assembled by the ribosome (the enzyme responsible for protein synthesis). The precise folding of a protein is dictated by its DNA sequence but researchers have not yet deciphered the rules for encoding structure by sequence ("the protein folding problem"). Addressing this problem is crucial to understanding how gene sequence variation translates into variation in protein and cell function. The delicate balance of forces which controls and guides the structural dynamics of the folding process is highly sensitive to environmental conditions inside the cell. The goal of this project is to synergistically apply cutting-edge methodologies, including single molecule spectroscopy, ultra-fast microfluidics mixing, photo-induced electron transfer, non-natural amino-acid labeling, mitochondrial protein transport, chemical peptide synthesis and simulation modeling using distributed and super-computing systems, to the study of protein folding under conditions that mimic the natural folding environment inside the living cell. The consortium of researchers will study the unfolded state of three different proteins in simple solutions (in-vitro) under a variety of conditions, and while the proteins are being made directly on the ribosome itself. By comparing such studies to protein folding experiments conducted within the crowded environment of the mitochondrial matrix (a mimic for the intracellular folding environment), this project seeks to understand the major differences between in-vitro and in-vivo folding environments and the effects of such differences on protein folding mechanisms. This project will have broad impacts on the field of cellular biology through the development of novel tools and methods as well as a general approach for studying complex biological processes on the molecular level. An outreach program will target the dissemination of these research tools to faculty and students from underrepresented institutions, and the enhancement of scientific and technological knowledge at the secondary education level. This project represents an interdisciplinary collaboration of researchers led by Shimon Weiss, at the University of California-Los Angeles with subawards to Stanford University (Vijay Pande), Texas A&M University (Arthur Johnson), University of California-Davis (Olgica Bakajin), Michigan State University (Lisa Lapidus) and Scripps Research Institute (Jeff Kelly). A large number of students and postdoctoral fellows will receive advanced training in conceptual and technical aspects of research at the interface of chemistry, biophysics, and simulation.