Exploration of target proteins for ubiquitination using fluorescence anisotropy measurement

本研究通过监测蛋白质的旋转运动来发现与泛素化相关蛋白相互作用的新因子。首先，我建立了实验程序来量化酵母膜脂的旋转运动。麦角甾醇生物合成缺陷导致膜刚性降低和脂质旋转运动增加。荧光素标记的模型蛋白的测量表明，这种技术是有效的蛋白质小于10 kDa。Sna3被鉴定为可能与Rsp5泛素连接酶相互作用的因子。在未来的研究中，我将使用本研究中使用的系统来研究Rsp5与Sna3之间的分子相互作用。

This study approaches the finding of novel factors that interact with ubiquitination-related proteins by monitoring the rotational motion of proteins. First, I established the experimental procedure to quantify the rotational motion of membrane lipids in yeast. The defect in ergosterol biosynthesis resulted in a decrease in membrane rigidity and an increase in the rotational lipid motion. The measurement of fluorescein-labeled model proteins suggests that this technique is valid for proteins of less than 10 kDa. Sna3 was identified as a factor that potentially interacted with Rsp5 ubiquitin ligase. In the future study, I investigate the molecular interaction between Rsp5 and Sna3 using the system employed in this study.