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对载脂蛋白B信使核糖核酸上组装的蛋白质复合物进行分析,以研究停泊序列依赖性核糖核酸编辑。

Analysis of protein complexes assembled on apolipoprotein B mRNA for mooring sequence-dependent RNA editing.

作者信息

Smith H C

机构信息

Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, New York 14642, USA.

出版信息

Methods. 1998 May;15(1):27-39. doi: 10.1006/meth.1998.0603.

Abstract

Apolipoprotein B (apoB) mRNA editing involves a cytidine-to-uridine transition at a select site catalyzed by a cytidine deaminase known as APOBEC-1. This enzyme cannot edit RNA alone but acquires site-specific editing capacity in the context of additional protein factors (auxiliary proteins). These proteins are currently hypothesized to assemble with APBEC-1 as a holoenzyme complex or editosome. Auxiliary proteins are the focus of ongoing research as they presumably serve important structural and regulatory roles in the editosome. The abilities of auxiliary proteins to interact with APOBEC-1 and apoB RNA and to promote RNA editing activity are important endpoints used as proof that proteins are functionally involved in apoB RNA editing. This article reviews the discovery of the editosome and provides detailed protocols for its isolation and subfractionation.

摘要

载脂蛋白B(apoB)信使核糖核酸(mRNA)编辑涉及一个特定位点的胞嘧啶到尿嘧啶的转变,该转变由一种名为载脂蛋白B信使核糖核酸编辑催化多肽1(APOBEC-1)的胞嘧啶脱氨酶催化。这种酶不能单独编辑核糖核酸(RNA),而是在其他蛋白质因子(辅助蛋白)的作用下获得位点特异性编辑能力。目前推测这些蛋白质与载脂蛋白B信使核糖核酸编辑催化多肽1(APBEC-1)组装成全酶复合物或编辑体。辅助蛋白是当前研究的重点,因为它们可能在编辑体中发挥重要的结构和调节作用。辅助蛋白与载脂蛋白B信使核糖核酸编辑催化多肽1(APOBEC-1)和载脂蛋白B(apoB)RNA相互作用以及促进RNA编辑活性的能力,是证明蛋白质在载脂蛋白B(apoB)RNA编辑中发挥功能作用的重要指标。本文综述了编辑体的发现,并提供了其分离和亚分级分离的详细方案。

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