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

FIBR：蛋白质如何在体外和活细胞的生理环境中折叠成其天然和功能结构？

• 批准号: 0623664
• 负责人: Shimon Weiss
• 金额: $ 455万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-10-01 至 2012-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0623664&HistoricalAwards=false
• 关键词: FIBR; How; Do; Proteins; Fold

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.

在所有活细胞中，蛋白质会自组装或折叠成精确的三维结构，具有由核糖体组装的线性多肽链的独特功能（负责蛋白质合成的酶）。 蛋白质的精确折叠取决于其DNA序列，但研究人员尚未通过序列编码结构的规则（“蛋白质折叠问题”）。 解决此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。 控制和指导折叠过程的结构动力学的力量的微妙平衡对细胞内部的环境条件高度敏感。 该项目的目的是协同采用尖端方法，包括单分子光谱，超快速的微流体混合，光诱导的电子传递，非北天然氨基酸标记，线粒体蛋白质的运输，化学肽合成和使用模拟模型，使用分布式和超级计算系统，研究在模仿活细胞内部自然折叠环境的条件下蛋白质折叠的研究。研究人员的联盟将在各种条件下研究三种不同蛋白的未折叠状态，而蛋白质直接在核糖体本身上制成。通过将此类研究与线粒体基质拥挤的环境中进行的蛋白质折叠实验进行比较（对细胞内折叠环境的模仿），该项目旨在理解视野内和体内折叠环境之间的主要差异以及这种影响蛋白质折叠机制的差异。 通过开发新工具和方法，该项目将对细胞生物学领域产生广泛的影响，以及在分子水平上研究复杂生物学过程的一般方法。 宣传计划将针对这些研究工具的传播给教师和代表性不足的机构的学生，并在中等教育水平上增强科学和技术知识。 该项目代表了由Shimon Weiss领导的研究人员的跨学科合作州立大学（Lisa Lapidus）和Scripps研究所（Jeff Kelly）。 许多学生和博士后研究员将在化学，生物物理学和仿真界面上接受研究的高级培训。