Hass Cathy S, Chen Ran, Wold Marc S
Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA, USA.
Methods Mol Biol. 2012;922:193-204. doi: 10.1007/978-1-62703-032-8_15.
Replication Protein A (RPA) is a single-strand DNA-binding protein that is found in all eukaryotes. RPA is subjected to multiple posttranslational modifications including serine- and threonine-phosphorylation, poly-ADP ribosylation, and SUMOylation. These modifications are believed to regulate RPA activity through modulating interactions with DNA and partner proteins. This article describes two methods used to detect posttranslational modified RPA: immunofluorescence and immmuoblotting.
复制蛋白A(RPA)是一种在所有真核生物中都能找到的单链DNA结合蛋白。RPA会经历多种翻译后修饰,包括丝氨酸和苏氨酸磷酸化、多聚ADP核糖基化以及小泛素样修饰蛋白化。据信这些修饰通过调节与DNA和伴侣蛋白的相互作用来调控RPA的活性。本文介绍了两种用于检测翻译后修饰的RPA的方法:免疫荧光法和免疫印迹法。