A NEW METHOD FOR PHOSPHORYLATION STUDIES

磷酸化研究的新方法

• 批准号: 6073693
• 负责人: Andrzej K Drukier
• 金额: $ 10.46万
• 依托单位: BIOTRACES, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6073693
• 关键词: calmodulin dependent protein kinase; fluorescence; method development; oligopeptides; phosphoproteins; phosphorylation; protein protein interaction; radiation detector

The goal of the proposed research is to develop novel methods for the detection of protein-protein interactions involving phosphoproteins and for intra and intermolecular distance measurements in various assemblies involving phosphoproteins - BioTraces' proprietary Multi Photon DetectionTM (MPDTM) system will be modified for the measurement of b-particle stimulated fluorescence (BSF) and the coincident detection of Cerenkov radiation excitation of fluorescence (CREF). The methods promise to provide a high sensitivity alternative to conventional fluorescence resonance energy transfer (FRET) techniques for monitoring the proximity of labeled biomoIecules. The methods will be tested on several model DNA and oligopeptide systems with the ultimate goal of applying them in studies of various CaM kinase II mediated phosphorylation cascades that are important for neuronal function. PROPOSED COMMERCIAL APPLICATIONS: The ability to characterize interactions among biomolecules and, in particular those involving phosphoproteins, is of immense importance to studies of the molecular basis of a variety of cellular functions, including various neuronal functions. Development of the methods proposed here will provide a set of tools for studying specific biomolecular interactions of importance for understanding physiological function and diseases at the cellular level.

拟议研究的目标是开发新方法，用于检测涉及磷蛋白的蛋白质-蛋白质相互作用，以及测量涉及磷蛋白的各种组件中的分子内和分子间距离 - BioTraces 专有的多光子检测TM (MPDTM) 系统将针对测量进行修改b 粒子受激荧光 (BSF) 和切伦科夫辐射激发荧光 (CREF) 的同时检测。该方法有望为传统荧光共振能量转移（FRET）技术提供高灵敏度替代方案，用于监测标记生物分子的接近度。这些方法将在几种模型 DNA 和寡肽系统上进行测试，最终目标是将它们应用于对神经元功能很重要的各种 CaM 激酶 II 介导的磷酸化级联的研究。拟议的商业应用：表征生物分子之间相互作用的能力，特别是那些涉及磷蛋白的相互作用，对于研究各种细胞功能（包括各种神经元功能）的分子基础非常重要。这里提出的方法的开发将为研究特定生物分子相互作用提供一套工具，这对于理解细胞水平的生理功能和疾病具有重要意义。