CAA: Chemical Synthesis of Affinity Tagged Flavonols

CAA：亲和标记黄酮醇的化学合成

• 批准号: 9722774
• 负责人: Loverine Taylor
• 金额: --
• 依托单位: Washington State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9722774&HistoricalAwards=false
• 关键词: CAA; Chemical; Synthesis; Affinity; Tagged

9722774 Taylor Using mutants that are unable to synthesize flavonols preliminary research has established an essential role of flavanols in reproduction. Although mutant, flavanol-deficient maize and petunia pollen is unable to germinate or produce a functional pollen tube, the defect can be corrected by the exogenous application of low concentrations of kaempferol, a flavonol aglycone to an in vitro suspension of pollen. This results in a rapid (2-5 min) pollen tube outgrowth that is indistinguishable from that of wild-type pollen. The rapid response of pollen to low doses of kaempferol indicates that this flavanol has a role in signal transduction. In order to understand how flavanols signal pollen germination it is essential to identify the proteins that interact with kaempferol. By using highly specific affinity probes it is feasible to identify the proteins that bind to kaempferol and thereby elucidate the downstream components of the signal transduction pathway. To gain new expertise in the synthesis and use of labeled flavanols as tools the following objectives will be accomplished as career advancement activities; 1, To synthesize and evaluate radiolabeled photoaffinity probes for flavonol (kaempferol) receptors. 2, To prepare kaempferol-polymer conjugates for affinity chromatography. 3, To make caged kaempferol to study the intracellular sites of action. 4, To adapt the existing yeast two-hybrid system to detect protein-protein interactions mediated by kaempferol. In order to become proficient in use of bioactive small molecule chemistry in biological research I will conduct sabbatical research and training in the laboratory of Dr. Thomas Wandless of the Department of Chemistry at Stanford University. As a consequence of the sabbatical research and training I will have expanded opportunities to investigate biological problems involving the signaling and regulation of pathways by small bioactive molecules. Many small molecules signal the onset of biological processes. How variou s molecules induce new physiological and developmental activity remains some of the most important biological processes that must be elucidated. Flavanols are essential to growth of pollen tubes and reproductive activity in plants. Mutants unable to produce flavanols are arrested in pollen growth that can be almost immediately restored with the exposure of the mutant pollen to low doses of the flavanol kaempferol. This grant will fund sabbatical training and research devoted toward developing tools and techniques that will allow the PI to investigate how flavanols interact with specific cellular proteins that result in signaling reproductive growth in pollen. ***

9722774 Taylor利用不能合成黄烷醇的突变体，初步研究已经确定了黄烷醇在生殖中的重要作用。虽然突变体，黄烷醇缺乏的玉米和矮牵牛花花粉不能发芽或产生功能性的花粉管，该缺陷可以通过外源应用低浓度的山奈酚，黄酮醇苷元在体外悬浮的花粉来纠正。这导致快速（2-5分钟）的花粉管生长，与野生型花粉无法区分。花粉对低剂量山奈酚的快速反应表明这种黄烷醇在信号转导中起作用。为了了解黄烷醇是如何发出花粉萌发信号的，识别与山奈酚相互作用的蛋白质至关重要。通过使用高度特异性的亲和探针，可以鉴定与山奈酚结合的蛋白质，从而阐明信号转导途径的下游组分。为了获得新的专业知识，在合成和使用标记的黄烷醇作为工具，以下目标将作为职业发展活动完成：1，合成和评价放射性标记的光亲和探针黄酮醇（山奈酚）受体。2、制备山奈酚-聚合物结合物用于亲和层析。3、制备笼状山萘酚，研究其在细胞内的作用部位。4、将现有的酵母双杂交系统用于检测山萘酚介导的蛋白质间相互作用。为了精通生物活性小分子化学在生物研究中的应用，我将在斯坦福大学化学系的托马斯旺德莱斯博士的实验室进行休假研究和培训。作为休假研究和培训的结果，我将有更多的机会来研究生物学问题，包括小生物活性分子的信号传导和途径调节。许多小分子是生物过程开始的信号。各种分子如何诱导新的生理和发育活动仍然是必须阐明的一些最重要的生物学过程。黄烷醇是植物花粉管生长和生殖活动所必需的。不能产生黄烷醇的突变体在花粉生长中被阻止，当突变体花粉暴露于低剂量的黄烷醇山萘酚时，花粉生长几乎可以立即恢复。这笔赠款将资助专门用于开发工具和技术的休假培训和研究，这些工具和技术将使PI能够研究黄烷醇如何与特定的细胞蛋白相互作用，从而导致花粉中的生殖生长信号。***