Aldol sensors for the rapid generation of tunable fluorescence by antibody catalysis
Aldol sensors for the rapid generation of tunable fluorescence by antibody catalysis
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DOI：
10.1073/pnas.95.26.15351
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发表时间：
1998-12-22
影响因子：
11.1
通讯作者：
Lerner, RA
中科院分区：
文献类型：
--
作者：
List, B;Barbas, CF;Lerner, RA
报道了醛缩酶抗体38C2的新型荧光逆醛缩醛底物的合成。这些底物可以被抗体醛缩酶有效和特异地处理，而不是由天然细胞酶处理。这些荧光底物和抗体醛缩酶一起提供了与活细胞兼容的报告基因系统。抗体醛缩酶的广泛范围允许处理一系列底物，这些底物可以被设计成允许在不同波长进行荧光监测。我们还在荧光蛋白标签中开发了以下概念。带有荧光标记的β-二酮被醛缩酶抗体共价结合合成羟醛传感器，而不是其他蛋白质。我们预计，与抗体融合的蛋白可以在活细胞内用几乎任何颜色的荧光团进行特异性和共价标记，从而提供绿色荧光蛋白融合的替代方案。
The synthesis of novel fluorogenic retro-aldol substrates for aldolase antibody 38C2 is described. These substrates are efficiently and specifically processed by antibody aldolases but not by natural cellular enzymes. Together, the fluorogenic substrates and antibody aldolases provide reporter gene systems that are compatible with living cells. The broad scope of the antibody aldolase allows for the processing of a range of substrates that can be designed to allow fluorescence monitoring at a variety of wavelengths. We also have developed the following concept in fluorescent protein tags. beta-Diketones bearing a fluorescent tag are bound Synthesis of the Aldol Sensors covalently by the aldolase antibody and not other proteins. We anticipate that proteins fused with the antibody can be tagged specifically and covalently within living cells with fluorophores of virtually any color, thereby providing an alternative to green fluorescent protein fusions.