Live-cell imaging of cellular proteins by a strain-promoted azide-alkyne cycloaddition.
Live-cell imaging of cellular proteins by a strain-promoted azide-alkyne cycloaddition.
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DOI：
10.1002/cbic.201000419
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发表时间：
2010-10-18
期刊：
影响因子：
3.2
通讯作者：
Tirrell, David A.
中科院分区：
文献类型：
--
作者：
Beatty, Kimberly E.;Fisk, John D.;Smart, Brian P.;Lu, Ying Ying;Szychowski, Janek;Hangauer, Matthew J.;Baskin, Jeremy M.;Bertozzi, Carolyn R.;Tirrell, David A.
蛋白质的荧光标记使得动态细胞过程可视化成为可能。标记通常是通过将目标蛋白融合到荧光蛋白（如绿色荧光蛋白（GFP））[1]或引导小分子荧光探针附着的短肽标签来完成的。[2]这些方法是研究预选蛋白质的理想方法。然而，由于这些蛋白质需要基因操作，它们不太适合研究整体细胞过程，如蛋白质合成或翻译后修饰。已经开发出策略，使蛋白质与酮，[3]叠氮化物，[4，5]或炔[6]功能基团在复杂的生物混合物中标记。在掺入反应性类似物后，已分离出糖基化、[7]磷酸化、[8]法尼基化、[9]和脂肪酰化[10]蛋白质。这些翻译后修饰是动态调节的，代谢标签使蛋白质组水平的分析成为可能，而这是遗传操作无法实现的。
Fluorescence labeling of proteins enables the visualization of dynamic cellular processes. Labeling is most often accomplished by fusing the protein of interest to a fluorescent protein, such as green fluorescent protein (GFP),[1] or to a short peptide tag that directs attachment of a small-molecule fluorescent probe.[2] These methods are ideal for the study of preselected proteins. However, because these proteins require genetic manipulation, they are not well suited to studies of global cellular processes, such as protein synthesis or posttranslational modification.Strategies have been developed to enable tagging of proteins with ketone,[3] azide,[4, 5] or alkyne [6] functional groups in complex biological mixtures. Glycosylated,[7] phosphorylated,[8] farnesylated,[9] and fatty-acylated [10] proteins have been isolated after the incorporation of reactive analogues. These post-translational modifications are regulated dynamically, and metabolic tagging has enabled proteome-level analyses that were not accessible by genetic manipulation.
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影响因子：
3.5
作者：
Kiick, KL;Weberskirch, R;Tirrell, DA
通讯作者：
Tirrell, DA
DOI：
10.1073/pnas.0506855103
发表时间：
2006-03-28
影响因子：
11.1
作者：
Dube, DH;Prescher, JA;Bertozzi, CR
通讯作者：
Bertozzi, CR
DOI：
10.1073/pnas.0707090104
发表时间：
2007-10-23
影响因子：
11.1
作者：
Baskin, Jeremy M.;Prescher, Jennifer A.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
2.7
作者：
Beatty KE;Tirrell DA
通讯作者：
Tirrell DA
DOI：
10.1073/pnas.0601637103
发表时间：
2006-06-20
影响因子：
11.1
作者：
Dieterich, Daniela C.;Link, A. James;Schuman, Erin M.
通讯作者：
Schuman, Erin M.