基于非天然氨基酸的蛋白质生物正交标记

doi:10.6023/A12050263

摘要/Abstract

生物正交化学反应正日益成为在活体内对生物大分子进行特异标记的一种有效方法. 最近涌现出的一个突出的例子是将金属钯催化的碳碳偶联反应这一在有机合成领域具有里程碑意义的反应拓展到生物大分子的标记上. 在活细胞上进行生物正交反应的一个先决条件是需要将参与这类反应的正交官能团特异地引入到目标生物大分子当中. 遗传密码子拓展技术是将多种生物正交活性基团引入到蛋白质当中的一种先进的手段; 最近发展出的基于吡咯赖氨酸氨酰合成酶和tRNA的体系能够将携带有生物正交官能团的非天然氨基酸有效地引入到原核生物、真核生物, 甚至是动物体内的蛋白质上. 在这一展望中, 我们首先介绍在生物正交反应和遗传密码子拓展这两个领域内的研究前沿与进展. 接着我们将讨论将这些新发展的研究工具, 尤其是基于钯催化的生物正交反应和基于吡咯赖氨酸氨酰合成酶的遗传密码子拓展技术, 应用于标记和修饰哺乳动物细胞蛋白质上的优势和诱人前景. 生物相兼容性更好的正交反应和更为灵活的非天然氨基酸引入技术必将有力地增强和拓宽人们在活细胞环境下特异操纵蛋白质的能力.

关键词: 蛋白质标记, 生物正交反应, 钯催化, 碳碳偶联, 非天然氨基酸

The bioorthogonal chemistry has become a valuable tool for specific labeling of biomolecules in living systems. One emerging example is to convert the palladium-mediated carbon-carbon bond formation reaction, a landmark reaction in organic synthesis, into the biocompatible reaction for biomolecule modifications. A prerequisite to enable such reactions to be executed on biological samples is to incorporate the corresponding bioorthogonal functional groups into target biomolecules. The genetic code expansion strategy is a leading technology on introducing diverse bioorthogonal functionalities into proteins, and the most recently developed pyrrolysine system can now allow unnatural amino acids (UAAs) bearing bioorthogonal handles into proteins in prokaryotic cells, eukaryotic cells and even in living animals. In this perspective, we will first introduce some of the forefronts of the bioorthognal reactions as well as the genetic code expansion strategy. We then will discuss the advantages and attractiveness of combining these newly emerged tools, Pd-mediated biomolecular labeling and pyrrolysine-based genetic code expansion system in particular, to specifically label and study proteins in living mammalian cells. More biocompatible reactions and more flexible UAA incorporation system will significantly strengthen and expand our ability to specifically manipulate proteins in their native cellular environment.

Key words: protein labeling, bioothorgonal, palladium catalyzed, cross coupling, unnatural amino acid

引用此文

李劼, 王杰, 陈鹏. 基于非天然氨基酸的蛋白质生物正交标记[J]. 化学学报, 2012, 70(13): 1439-1445.

Li Jie, Wang Jie, Chen Peng R.. Unnatural Amino Acid Mediated Protein Bioorthogonal Labeling[J]. Acta Chimica Sinica, 2012, 70(13): 1439-1445.

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