活细胞中用合成荧光团对融合蛋白进行标记。
Labeling of fusion proteins with synthetic fluorophores in live cells.
作者信息
Keppler Antje, Pick Horst, Arrivoli Claudio, Vogel Horst, Johnsson Kai
机构信息
Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
出版信息
Proc Natl Acad Sci U S A. 2004 Jul 6;101(27):9955-9. doi: 10.1073/pnas.0401923101. Epub 2004 Jun 28.
Abstract
A general approach for the sequential labeling of fusion proteins of O(6)-alkylguanine-DNA alkyltransferase (AGT) with different fluorophores in mammalian cells is presented. AGT fusion proteins with different localizations in the cell can be labeled specifically with different fluorophores, and the fluorescence labeling can be used for applications such as multicolor analysis of dynamic processes and fluorescence resonance energy transfer measurements. The facile access to a variety of different AGT substrates as well as the specificity of the labeling reaction should make the approach an important tool to study protein function in live cells.
摘要
本文介绍了一种在哺乳动物细胞中对O(6)-烷基鸟嘌呤-DNA烷基转移酶（AGT）融合蛋白进行不同荧光团顺序标记的通用方法。细胞中具有不同定位的AGT融合蛋白可以用不同的荧光团进行特异性标记，并且荧光标记可用于动态过程的多色分析和荧光共振能量转移测量等应用。能够轻松获得多种不同的AGT底物以及标记反应的特异性，应使该方法成为研究活细胞中蛋白质功能的重要工具。
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