MUTIDIMENSIONAL DIFFUSION EXPERIMENTS IN A CELL-FREE SYSTEM

无细胞系统中的多维扩散实验

• 批准号: 7420534
• 负责人: Silvia Cavagnero
• 金额: $ 0.6万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-20 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420534
• 关键词: carbohydrates; cell free system; diffusion; proteins; quinine; small molecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Diffusion-ordered spectroscopy (DOSY) is an effective tool to spectroscopically discriminate small molecules within complex mixtures based on differences in their diffusion coefficient in solution.This method has typically been applied to mixtures of relatively small molecules (quinine, camphene, geraniol, carbohydrates, etc.). No method is currently available to discriminate large polypeptides and proteins from smaller molecules.We have developed the DOSY-HSQC pulse sequence for the straightforward spectroscopic separation of resonances due to proteins from those due to small molecules.This method can be applied to the spectroscopic analysis of complex biological media, including proteolytic mixtures, living cells, and cell-free systems containing overexpressed isotopically labeled proteins

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。扩散有序光谱 (DOSY) 是一种根据溶液中扩散系数的差异对复杂混合物中的小分子进行光谱区分的有效工具。该方法通常应用于相对小分子（奎宁、莰烯、香叶醇、碳水化合物等）的混合物。目前还没有方法可以区分大的多肽和蛋白质与较小的分子。我们开发了 DOSY-HSQC 脉冲序列，用于直接光谱分离蛋白质引起的共振和小分子引起的共振。该方法可应用于复杂生物介质的光谱分析，包括蛋白水解混合物、活细胞和含有过表达同位素的无细胞系统 标记蛋白质