权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development and Applications of Bioorthogonal Chemistry: Administrative Supplement for Equipment

生物正交化学的发展与应用：设备管理补充

基本信息

批准号：
10581256
负责人：
Qing Lin
金额：
$ 7.44万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2023-12-31
项目状态：
已结题

项目摘要

Development and Applications of Bioorthogonal Chemistry: Administrative Supplement for Equipment ABSTRACT We have a long-standing interest in developing reactivity-based chemical tools to address significant biological problems that are difficult to solve using conventional molecular biology techniques. In our MIRA application, we plan to continue our studies of orthogonal chemical reactivity at the chemistry-biology interface and pursue the following two related projects. In Project 1, we will construct the FRET-based biosensors of GLP-1R and GCGR using bioorthogonal chemistry techniques. These biosensors will then be employed to probe the conformational dynamics during the ligand-induced receptor activation and signaling in live cells. A new set of fluorescence 'turn-on' reagents will be designed for bioorthogonal labeling of the intracellular loop 3 of GLP-1R and GCGR to allow single-cell intra- and inter-molecular FRET analysis of receptor dynamics in live cells. In Project 2, we will develop a genetically encoded chemical crosslinker containing an alkyne group and apply this chemical crosslinker to map the time-dependent GLP-1R and -arrestins interactomes by mass spectrometry in response to ligand stimulation. We expect these studies to validate new bioorthogonal tools for real-time monitoring of protein conformation and protein-protein interactions in live cells. Also, we will gain novel insights into the GLP-1R and GCGR activation dynamics and structural basis of biased signaling that are crucial for the development of targeted therapies for the treatment of diabetes and obesity. This Administrative Supplement requests the acquisition of an Agilent 1260 Infinity II LC System with Diode Array Detector. This system will help us characterize the bioorthogonal reagents' reaction kinetics and selectivity at higher throughput and greater accuracy.

生物正交化学的发展与应用--设备管理补充摘要我们长期以来一直致力于开发基于反应性的化学工具，以解决重大的生物问题。这些问题很难用传统的分子生物学技术来解决。在我们的MIRA应用程序中，我们计划继续研究化学-生物学界面上的正交化学反应性，以下两个相关项目。在项目1中，我们将构建基于FRET的GLP-1 R生物传感器，使用生物正交化学技术的GCGR。这些生物传感器将被用来探测在活细胞中配体诱导的受体活化和信号传导期间的构象动力学。一套新的将设计荧光“开启”试剂用于GLP-1 R细胞内环3的生物正交标记和GCGR，以允许活细胞中受体动力学的单细胞内和分子间FRET分析。在项目二，我们将开发一种含有炔基的基因编码化学交联剂，并应用于该化学交联剂用于通过质量绘制时间依赖性GLP-1 R和β-抑制蛋白相互作用组响应于配体刺激的光谱分析。我们希望这些研究能够验证新的生物正交工具，实时监测活细胞中的蛋白质构象和蛋白质-蛋白质相互作用。同时，我们将获得新的见解GLP-1 R和GCGR激活动力学和结构基础的偏见信号，这对于开发治疗糖尿病和肥胖症的靶向疗法至关重要。这行政补充申请要求采购Agilent 1260 Infinity II LC系统（带二极管）阵列检测器。该系统将帮助我们表征生物正交试剂的反应动力学，选择性更高的吞吐量和更高的精度。