Collaborative Research-Lanthanide Binding Tags: Chemical Tools for Investigating Protein Structure and Function

协作研究-镧系元素结合标签：研究蛋白质结构和功能的化学工具

• 批准号: 0744415
• 负责人: Karen Allen
• 金额: $ 79.12万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0744415&HistoricalAwards=false
• 关键词: Collaborative; Research; Lanthanide; Binding; Tags

Intellectual Merit: Lanthanide binding tags (LBTs) are small peptides (15-20 residues) that impart on the protein to which they are attached specific and tight binding of lanthanide ions. LBTs permit the use of applications that rely upon the photophysical properties of the bound lanthanides. These sequences have been integrated into many target proteins and tested for applications in nuclear magnetic resonance, X-ray crystallography, and luminescence. In the next phase of research, the LBT technology will be enlisted for addressing challenging problems of biological significance, moving from the test tube to the cell. The new studies will exploit the unique properties of the LBTs and the bound lanthanides. Namely, lanthanides luminescence properties can eliminate the high background often complicating measurements in biological systems using current technologies for locating proteins and measuring protein motion. The LBT capabilities will be extended by modification of the chemistry to impart properties with greater number of potential application. There are four parts to the project: 1) The LBTs will be used to study dynamic changes in protein structure using the property of luminescence resonance energy transfer in the important cell signaling calcineurin A/calcineurin B complex. 2) These new LBTs will be used to increase understanding of cell-cell signaling and cellular trafficking. 3) The LBT will be developed as a novel gadolinium contrast agent for use in magnetic resonance imaging. 4) The use of LBTs in determining protein crystal structures will be extended to the challenging problem of solution structure of proteins embedded in cell membranes. The LBTs will provide multiple features for the solution of these difficult structures. The dissemination of the information gained from these studies will allow the broad application of LBTs by the research communities. During the development of the tags, high school students, graduate students and postdoctoral associates will been trained in an integrated and collaborative research environment spanning multiple disciplines. These studies will provide the researchers with the foundation needed to address a broad range of problems at the interface of Biology, Chemistry and Physics. Broader Impact of the Research. During the project period, use of the LBTs will be extended in applications of biological importance, providing the scientific community with new LBTs and defined protocols for the application of the tags to a wide variety of problems including the study of protein dynamics, protein signaling and trafficking, cellular environment reporting, and the structure of integral membrane proteins. During the development of the tags, students and postdoctoral associates will be trained in an integrated and collaborative research environment spanning skill sets in molecular biology, fluorescence, protein chemistry, MRI, and X-ray crystallography at multiple institutions. These studies will provide the researchers with the foundation needed to address a broad range of problems at the interface of Biology, Chemistry and Physics. Undergraduate students will have the opportunity to be trained under the auspices of the UROP (undergraduate research opportunities) programs at MIT and Boston University School of Medicine as well as High School students through the BU CityLab program. Both MIT and Boston University have aggressive policies and procedures in action for the recruitment and retention of under-represented minorities at the undergraduate and graduate levels.

智力优势：镧系元素结合标签（LBT）是小肽（15-20个残基），其赋予与其连接的蛋白质镧系元素离子的特异性和紧密结合。LBT允许使用依赖于结合的镧系元素的电子物理性质的应用。这些序列已被整合到许多靶蛋白中，并在核磁共振、X射线晶体学和发光中进行了应用测试。在下一阶段的研究中，LBT技术将被用于解决具有生物学意义的挑战性问题，从试管转移到细胞。新的研究将利用LBT和结合镧系元素的独特性质。也就是说，镧系元素发光特性可以消除高背景，这通常使使用当前技术定位蛋白质和测量蛋白质运动的生物系统中的测量复杂化。LBT的能力将通过化学改性来扩展，以赋予具有更多潜在应用的特性。本课题主要包括四个部分：1）利用钙调神经磷酸酶A/钙调神经磷酸酶B复合物的发光共振能量转移特性，研究蛋白质结构的动态变化。2)这些新的LBT将用于增加对细胞间信号传导和细胞运输的理解。3)LBT将被开发为一种新型的钆造影剂，用于磁共振成像。4)LBT在确定蛋白质晶体结构中的应用将扩展到嵌入细胞膜的蛋白质的溶液结构的挑战性问题。LBT将为这些困难结构的解决方案提供多种功能。传播从这些研究中获得的信息将使研究界能够广泛应用LBT。在标签的开发过程中，高中生，研究生和博士后助理将在跨越多个学科的综合和协作研究环境中接受培训。这些研究将为研究人员提供解决生物学，化学和物理学界面上广泛问题所需的基础。研究的更广泛影响。在项目期间，LBT的使用将扩展到具有生物学重要性的应用中，为科学界提供新的LBT和定义的协议，用于将标签应用于各种各样的问题，包括蛋白质动力学，蛋白质信号传导和运输，细胞环境报告以及完整膜蛋白的结构。在标签的开发过程中，学生和博士后将在多个机构的分子生物学，荧光，蛋白质化学，MRI和X射线晶体学的综合和协作研究环境中接受培训。这些研究将为研究人员提供解决生物学，化学和物理学界面上广泛问题所需的基础。本科生将有机会在麻省理工学院和波士顿大学医学院的UROP（本科生研究机会）项目的主持下接受培训，高中生也有机会通过BU CityLab项目接受培训。麻省理工学院和波士顿大学都有积极的政策和程序，在本科和研究生阶段招聘和保留代表性不足的少数民族。