Coiled-coil Modules for Molecular Engineering and Synthetic Biology

用于分子工程和合成生物学的卷曲螺旋模块

• 批准号: 0950233
• 负责人: Amy Keating
• 金额: $ 86万
• 依托单位: Massachusetts Institute of Technology
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0950233&HistoricalAwards=false
• 关键词: Coiled; coil; Modules; Molecular; Engineering

Intellectual Merit: Understanding how amino-acid sequences determine the structure and function of proteins provides an opportunity to engineer molecular tools at the nanoscale level. The alpha helical coiled coil is a simple protein structure made up of two or more chains that associate into a rod-like super-helix. Several decades of research have provided insights into the principles that determine whether two protein chains can associate to form a coiled coil. This process can now be controlled, to a certain extent, by manipulating protein sequence using rational molecular design techniques. This project aims to exploit current understanding of coiled-coil sequence-structure relationships, along with powerful molecular engineering techniques, to develop reagents suitable for building new materials and re-engineering cellular circuits using coiled coil motifs. The goal is to generate a list of fully described molecular parts that will allow other scientists to use these as modular units in bioengineering projects. The research builds on the prior identification of 23 synthetic proteins that participate in a variety of complex interaction patterns. Goals for the project are to (1) test whether these peptides, previously characterized in solution in the laboratory, maintain their interaction properties inside bacteria and yeast, (2) refine and tune the interaction strengths and specificities of these 23 proteins through rational molecular design, (3) explore how the modular coiled-coil units can be used to alter information processing in laboratory yeast, specifically by modifying MAP-kinase signaling, and (4) explore the use of modular coiled-coil proteins for constructing artificial DNA-binding proteins. Success in these areas will pave the way for broad use of synthetic coiled coils in molecular engineering applications. As modular design blocks adapted from the natural world, coiled coils have the potential to advance materials science, provide new capabilities for engineering microbial cells, and interface with nanoscale inorganic materials, enabling advanced information processing and smart devices.Broader Impacts: Beyond the scientific impact, this project will enhance integrated educational/research opportunities for a diverse group of young scientists. The proposed work will be carried out in the Keating laboratory in the MIT biology department, where education and research activities are tightly coupled. The Keating research group of 12 individuals is composed of graduate students, postdoctoral scientists and undergraduate students. Laboratory members with diverse scientific, economic, ethnic and cultural backgrounds are involved in all group activities, and Dr. Keating additionally participates in mentoring activities for women and under-represented students in science. Undergraduate students will be recruited to work on this project, with summer invitations extended particularly to individuals not likely otherwise to be exposed to modern research. Protein engineering and synthetic biology are appealing research subjects for undergraduates, because these provide the opportunity to rationally construct something entirely new in the molecular world. This will enhance the spirit of exploration and discovery that is critical for engaging and retaining young scientists.

智力优势：了解氨基酸序列如何决定蛋白质的结构和功能，为在纳米级水平上设计分子工具提供了机会。α螺旋卷曲螺旋是一种简单的蛋白质结构，由两条或更多条链组成，这些链结合成棒状超螺旋。几十年的研究已经提供了关于决定两条蛋白质链是否可以结合形成卷曲螺旋的原理的见解。这个过程现在可以控制，在一定程度上，通过操纵蛋白质序列使用合理的分子设计技术。本项目旨在利用目前对卷曲螺旋序列-结构关系的理解，沿着强大的分子工程技术，开发适用于使用卷曲螺旋基序构建新材料和重新设计细胞电路的试剂。目标是生成一个完整描述的分子部分列表，允许其他科学家在生物工程项目中将其作为模块单元使用。这项研究建立在先前鉴定的23种参与各种复杂相互作用模式的合成蛋白质的基础上。该项目的目标是（1）测试这些肽，以前在实验室中的溶液中表征，是否保持它们在细菌和酵母中的相互作用特性，（2）通过合理的分子设计改进和调整这23种蛋白质的相互作用强度和特异性，（3）探索模块化卷曲螺旋单元如何用于改变实验室酵母中的信息处理，特别是通过修饰MAP-激酶信号传导，和（4）探索模块化卷曲螺旋蛋白用于构建人工DNA结合蛋白的用途。这些领域的成功将为合成卷曲螺旋在分子工程应用中的广泛应用铺平道路。作为从自然界改造而来的模块化设计模块，螺旋线圈有可能推进材料科学的发展，为工程微生物细胞提供新的能力，并与纳米无机材料相结合，实现先进的信息处理和智能设备。更广泛的影响：除了科学影响，该项目还将为多样化的年轻科学家群体提供综合教育/研究机会。拟议的工作将在麻省理工学院生物系的基廷实验室进行，那里的教育和研究活动紧密结合。基廷研究小组由12名研究生、博士后科学家和本科生组成。具有不同科学、经济、种族和文化背景的实验室成员参与了所有小组活动，基廷博士还参加了为妇女和科学领域代表性不足的学生举办的辅导活动。本科生将被招募到这个项目中工作，特别是对那些不太可能接触现代研究的人发出夏季邀请。蛋白质工程和合成生物学对本科生来说是很有吸引力的研究课题，因为它们提供了在分子世界中合理构建全新事物的机会。这将增强探索和发现的精神，这对吸引和留住年轻科学家至关重要。