A NEW METHOD FOR PHOSPHORYLATION STUDIES

提出的研究目标是开发新的方法来检测涉及磷蛋白的蛋白质-蛋白质相互作用，以及在涉及磷蛋白的各种组件中进行分子内和分子间距离测量- BioTraces专有的多光子检测tm （MPDTM）系统将被修改用于测量b粒子受激荧光（BSF）和切伦科夫辐射激发荧光（CREF）的同步检测。这些方法有望提供一种高灵敏度替代传统的荧光共振能量转移（FRET）技术，用于监测标记生物分子的接近性。这些方法将在几个模型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重要的各种CaM激酶II介导的磷酸化级联反应的研究中。建议的商业应用：表征生物分子之间相互作用的能力，特别是涉及磷酸化蛋白的相互作用，对于研究各种细胞功能的分子基础，包括各种神经元功能，是非常重要的。本文提出的方法的发展将为研究特定的生物分子相互作用提供一套工具，这对于在细胞水平上理解生理功能和疾病具有重要意义。

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.