一种用于选择性分离多聚泛素化蛋白的平行亲和纯化方法。

A parallel affinity purification method for selective isolation of polyubiquitinated proteins.

作者信息

Ota Kazuhisa, Kito Keiji, Iemura Shun-ichiro, Natsume Tohru, Ito Takashi

机构信息

Department of Computational Biology, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan.

出版信息

Proteomics. 2008 Aug;8(15):3004-7. doi: 10.1002/pmic.200800271.

DOI:10.1002/pmic.200800271

PMID:18615433

Abstract

We developed a parallel affinity purification (PAP) procedure, in which ubiquitinated proteins are purified from the cells that coexpress two affinity-tagged ubiquitins by sequential use of affinity chromatography specific to each tag. In contrast with previous procedures using a single affinity-tagged ubiquitin, the PAP eliminates highly abundant ubiquitin monomers and monoubiquitinated proteins to selectively enrich proteins bearing both affinity-tags, or poly- and multiubiquitinated proteins. Accordingly, it would serve as a powerful method to facilitate mass-spectrometric identification of ubiquitinated proteins.

摘要

我们开发了一种平行亲和纯化（PAP）方法，该方法通过依次使用针对每个标签的亲和色谱，从共表达两种带有亲和标签的泛素的细胞中纯化泛素化蛋白。与先前使用单一带有亲和标签的泛素的方法不同，PAP 消除了高度丰富的泛素单体和单泛素化蛋白，以选择性富集带有两个亲和标签的蛋白或多聚泛素化和多泛素化蛋白。因此，它将成为促进泛素化蛋白质谱鉴定的有力方法。