CONFORMATIONS OF LIGHT-SENSITIVE PROTEINS

光敏蛋白的构象

基本信息

批准号：
7954904
负责人：
Tobin R Sosnick
金额：
$ 0.66万
依托单位：
ILLINOIS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2009-12-31
项目状态：
已结题

项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A major aim in structural biology is to design new proteins and control their function, including novel protein-to-protein signalling. The modular nature of proteins makes such engineering possible, but mechanisms by which component parts can be connected have not been fully elucidated. We have developed a simple method to couple the functions of two proteins, which we demonstrate here with light-triggered DNA binding. When illuminated, this fusion of a photoactive LOV domain and bacterial trp repressor selectively binds operator DNA upon illumination. This work suggests new ways of reprogramming cellular behaviors such as metabolism, morphology, and cell death. We are interested in investigating how large scale conformational changes are involved in the propagation of signals in our designed protein. Recent work has shown that some, and perhaps many, LOV domains respond to light with rather large structural changes. Our design strategy aims to take advantage of these changes, and measuring their magnitude and how they correlate with activity is an important means of validating our results. The SAXS experiments we have undertaken at BioCAT have been a critical part of this work. In some respects they have confirmed our expectations and in others they have challenged them and led us to refine our model. Paper accepted at PNAS pending minor revision: Light-activated DNA binding in a designed allosteric protein. Devin Strickland, Keith Moffat, and Tobin R. Sosnick

该副本是利用众多研究子项目之一由NIH/NCRR资助的中心赠款提供的资源。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构是对于中心，这不一定是调查员的机构。结构生物学的主要目的是设计新蛋白质并控制其功能，包括新型蛋白质对蛋白质信号。蛋白质的模块化性质使这种工程成为可能，但是可以连接组件部分的机制尚未完全阐明。我们已经开发了一种简单的方法来培养两种蛋白质的功能，我们在这里以光触发的DNA结合在此处证明。当照明时，光活性LOV结构域和细菌TRP阻遏物的这种融合会在照明时选择性地结合操作员DNA。这项工作提出了重新编程细胞行为（例如代谢，形态和细胞死亡）的新方法。我们有兴趣研究我们设计蛋白质中信号的传播涉及大规模构象的变化。最近的工作表明，有些（也许还有许多LOV领域）会随着结构上的巨大变化做出响应。我们的设计策略旨在利用这些变化，衡量它们的幅度以及它们与活动的相关性是验证我们结果的重要手段。我们在BioCat进行的SAXS实验一直是这项工作的关键部分。在某些方面，他们确认了我们的期望，在其他方面，他们对他们挑战，并使我们改善了模型。在PNAS上接受的纸张即将进行较小的修订：在设计的变构蛋白中，光激活的DNA结合。 Devin Strickland，Keith Moffat和Tobin R. Sosnick