Addition of a photocrosslinking amino acid to the genetic code of Escherichia coli
Addition of a photocrosslinking amino acid to the genetic code of Escherichia coli
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DOI：
10.1073/pnas.172226299
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发表时间：
2002-08-20
影响因子：
11.1
通讯作者：
Schultz, PG
中科院分区：
文献类型：
--
作者：
Chin, JW;Martin, AB;Schultz, PG
二苯甲酮是生物学中最有用的光交联剂之一。我们已经进化出一个正交的氨基酰基-tRNA合成酶/tRNA对，使对苯甲酰- l-苯丙氨酸在大肠杆菌中响应琥珀色密码子TAG的体内结合到蛋白质中成为可能。该非天然氨基酸以高翻译效率和保真度结合到二聚体蛋白谷胱甘肽s -转移酶中。辐照导致蛋白质亚基的有效交联（约50%）。这种方法可能被证明是有用的发现和确定蛋白质相互作用在体外和体内。
Benzophenones are among the most useful photocrosslinking agents in biology. We have evolved an orthogonal aminoacyl-tRNA synthetase/tRNA pair that makes possible the in vivo incorporation of p-benzoyl-L-phenylalanine into proteins in Escherichia coli in response to the amber codon, TAG. This unnatural amino acid was incorporated with high translational efficiency and fidelity into the dimeric protein glutathione S-transferase. Irradiation resulted in efficient crosslinking (>50%) of the protein subunits. This methodology may prove useful for discovering and defining protein interactions in vitro and in vivo.