Luminometer for the discovery of novel bioactive compounds

用于发现新型生物活性化合物的发光计

• 批准号: 344621-2007
• 负责人: Aluko, Rotimi
• 金额: $ 1.1万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2006
• 资助国家: 加拿大
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=325559
• 关键词: Luminometer; discovery; novel; bioactive; compounds

Calmodulin (CaM)-dependent 3',5'-cyclic nucleotide phosphodiesterase (CaM-PDE) is a cellular enzyme that hydrolyzes cyclic adenosine monophosphate (cAMP) to form 5'-adenosine monophosphate. In normal human cells, elevated levels of cAMP is required to inhibit excessive proliferation and to induce normal cell death. However, in diseased states such as cancer, there is excessive levels of CaM-PDE, which decreases cAMP concentrations and disrupts the normal cell death that is required to check excessive cellular proliferation. Current evidence also indicates that excessive levels of CaM-PDE play significant roles in several pathobiologic processes leading to kidney fibrosis and hypertrophy of the heart. The aim of this proposal is to purchase a luminometer that will enhance discovery and identification of CaM-binding peptides that inhibit the activity of CaM-PDE with potential application as therapeutic agents against chronic diseases such as cancer and kidney fibrosis. The luminometer is the most sensitive and accurate instrument for measuring the light output during CaM-PDE reaction that has been coupled to the luciferin-luciferase system. The instrument also provides for the most rapid form of analysis of CaM-PDE activity. Therefore, the instrument will enhance the screening of several hundreds of potential peptide inhibitors with high accuracy and reliability over a short period of time. However, there is no luminometer in the Department of Human Nutritional Sciences at the University of Manitoba. Results provided by the instrument will identify bioactive peptides that can be used to formulate functional foods and nutraceuticals, in addition to providing fundamental information on the structure and activity relationships of the CaM-binding peptides. Technology developed from this work will facilitate value-added utilization of Canadian-grown crops such as peas, flaxseed, soybean with economic returns to the Canadian economy. Availability of the instrument will enhance interdisciplinary training of highly qualified personnel such as postdoctoral fellows, graduate and undergraduate students in the areas of food science and life sciences related to human health.

钙调素依赖的3‘，5’-环核苷酸磷酸二酯酶(CaM-PDE)是一种能将环腺苷一磷酸(CAMP)水解成5‘-腺苷一磷酸的细胞酶。在正常人类细胞中，cAMP水平升高是抑制过度增殖和诱导正常细胞死亡所必需的。然而，在癌症等疾病状态下，CaM-PDE水平过高，这会降低cAMP浓度，并扰乱正常的细胞死亡，而正常细胞死亡是抑制细胞过度增殖所必需的。目前的证据还表明，过高的CaM-PDE水平在导致肾脏纤维化和心脏肥大的几个病理生物学过程中发挥着重要作用。这项提议的目的是购买一种发光计，该发光计将加强对抑制CaM-PDE活性的CaM结合肽的发现和鉴定，并有可能用作治疗癌症和肾脏纤维化等慢性疾病的药物。发光计是测量CaM-PDE反应过程中光输出的最灵敏和准确的仪器，该反应已耦合到荧光素-荧光素酶系统。该仪器还提供了CaM-PDE活性的最快速分析形式。因此，该仪器将在短时间内提高数百种潜在多肽抑制剂的筛选精度和可靠性。然而，马尼托巴大学人类营养科学系没有发光计。该仪器提供的结果将鉴定可用于配制功能食品和营养食品的生物活性多肽，此外还将提供有关CaM结合多肽的结构和活性关系的基本信息。这项工作开发的技术将促进加拿大种植的作物(如豌豆、亚麻籽、大豆)的增值利用，为加拿大经济带来经济回报。该仪器的提供将加强对高素质人员的跨学科培训，如与人类健康有关的食品科学和生命科学领域的博士后研究员、研究生和本科生。