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

智能优点：镧系元素结合标签(LBT)是一种小肽(15-20个残基)，赋予它们所附着的蛋白质对稀土离子的特定和紧密的结合。LBT允许使用依赖于结合的稀土元素的光物理性质的应用。这些序列已经整合到许多靶蛋白中，并在核磁共振、X射线结晶学和发光方面进行了应用测试。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LBTS和结合的稀土元素的独特性质。也就是说，使用当前用于定位蛋白质和测量蛋白质运动的技术，镧系元素的发光特性可以消除生物系统中经常使测量复杂化的高背景。LBT的能力将通过化学修饰来扩展，从而赋予性能更多的潜在应用。该项目包括四个部分：1)利用钙调神经磷酸酶A/B复合体的发光共振能量传递特性，利用LBTS研究蛋白质结构的动态变化。2)这些新的LBT将被用来增加对细胞间信号和细胞运输的理解。3)LBT将被开发为一种用于磁共振成像的新型Gd造影剂。4)LBTS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溶液结构这一具有挑战性的问题。LBTS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LBTS。在TAG的开发过程中，高中生、研究生和博士后助理将在跨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所需的基础，以解决生物、化学和物理之间的广泛问题。研究的更广泛影响。在项目期间，LBTS的使用将扩展到具有生物学意义的应用中，为科学界提供新的LBTS和定义的协议，用于将标签应用于各种问题，包括蛋白质动力学、蛋白质信号和运输、细胞环境报告和完整膜蛋白质的结构的研究。在TAG的开发过程中，学生和博士后将在多个机构的综合和协作研究环境中接受培训，这些研究包括分子生物学、荧光、蛋白质化学、核磁共振和X射线结晶学方面的技能。这些研究将为研究人员提供所需的基础，以解决生物、化学和物理之间的广泛问题。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)计划下接受培训，而高中学生将有机会通过波士顿大学城市实验室计划接受培训。麻省理工学院和波士顿大学都有积极的政策和程序，在本科生和研究生一级招聘和留住代表不足的少数族裔。

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.