Covalent Protein Labeling Based on Noncatalytic β-Lactamase and a Designed FRET Substrate
Covalent Protein Labeling Based on Noncatalytic β-Lactamase and a Designed FRET Substrate
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DOI：
10.1021/ja8082285
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发表时间：
2009-04-15
影响因子：
15
通讯作者：
Kikuchi, Kazuya
中科院分区：
文献类型：
--
作者：
Mizukami, Shin;Watanabe, Shuji;Kikuchi, Kazuya
用小分子标记蛋白质的技术引起了许多生命科学家的注意。我们开发了一种新的蛋白质标记系统，该系统结合了一种转基因的、非催化的b -内酰胺酶变体和基于特定机制的荧光探针。标记蛋白和标记探针的合理设计使氟化物的结合具有高度特异性。凝胶电泳、质谱、荧光光谱和荧光显微成像证实了我们方法的可行性。满足特异性和致氟性双重标准的标记技术很少有报道。因此，这种方法可以成为生命科学领域广泛使用的研究工具。
Techniques for labeling proteins with small molecules have attracted the attention of many life scientists. We have developed a novel protein labeling system that combines a genetically modified, noncatalytic B-lactamase variant and specific mechanism-based fluorescent probes. Rational design of the tag protein and the labeling probes enables highly specific incorporation of the fluorogen. The feasibility of our approach was confirmed by gel electrophoresis, mass spectrometry, fluorescence spectroscopy, and fluorescence microscopic imaging. Labeling techniques that satisfy the dual criteria of specificity and fluorogenicity have rarely been reported. As a consequence, this method could be a broadly useful research tool in the field of life science.