Position-specific incorporation of a highly photodurable and blue-laser excitable fluorescent amino acid into proteins for fluorescence sensing
Position-specific incorporation of a highly photodurable and blue-laser excitable fluorescent amino acid into proteins for fluorescence sensing
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DOI：
10.1016/j.bmc.2005.03.014
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发表时间：
2005-05-16
影响因子：
3.5
通讯作者：
Sisido, M
中科院分区：
文献类型：
--
作者：
Hamada, H;Kameshima, N;Sisido, M
利用4碱基密码子/反密码子策略，将新的荧光氨基酸L-2-吖啶基丙氨酸掺入蛋白质的特定位置。掺入的效率足够高，可以获得足够数量的突变体。当在蓝色激光的波长下被激发时，吖啶酮基团是高度荧光的，与通常用于生物分析的传统荧光团相比，它具有很高的耐光性。荧光强度对环境极性的微小变化很敏感。当非天然氨基酸被结合到链霉亲和素的特定位置时，突变蛋白作为生物素的荧光传感器工作。同样，当氨基酸被掺入到单链抗体中时，突变蛋白对抗原分子有灵敏的反应。由于具有高的掺入效率、高的耐光性、对蓝光的可激活性以及对环境的高度敏感性等特点，吖啶基丙氨酸被多种抗体、受体和酶结合到特定的位置，成为一种很有前途的荧光氨基酸，用于检测小分子。(C)2005爱思唯尔有限公司。保留所有权利。
A new fluorescent amino acid, L-2-acridonylalanine, was incorporated into proteins at specific positions using 4-base codon/anticodon strategy. The efficiency of the incorporation was high enough to obtain enough quantities of the mutants. The acridonyl group was highly fluorescent when it was excited at the wavelengths of blue-lasers and was highly photodurable compared with conventional fluorophores often used for biological analyses. The fluorescence intensity was sensitive to small changes in the polarity of the environment. When the nonnatural amino acid was incorporated into specific positions of streptavidin, the mutant protein worked as a fluorescent sensor to biotin. Similarly, when the amino acid was incorporated into came] single-chain antibody, the mutant protein sensitively responded to the antigen molecule. The high incorporation efficiency, the high photodurability, the excitability with blue-lasers, and high sensitivity to the environment make the acridonylalanine as the promising fluorescent amino acid for sensing small molecules when incorporated into specific positions of various antibodies, receptors, and enzymes. (c) 2005 Elsevier Ltd. All rights reserved.