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用3′末端氧化的精氨酰tRNA共价修饰大肠杆菌精氨酰tRNA合成酶,酶的ATP-PPi交换活力和氨酰化活力完全丧失。用SDS-PAGE分析酶与tRNA(?)形成的共价复合物发现:伴随着酶的失活,形成了两种形式明显不同的ArgRS-tRNA(?)复合物,它们对应于凝胶上两条迁移率不同的大分子条带。这一结果与其它合成酶亲和标记的结果不同。ArgRS-tRNA(?)经RNase处理后,ATP-PPi交换活力和氨酰化活力均有部分恢复(小于15％)。在整个标记过程中,酶的两个活力是紧密相关联的。
The arginyl tRNA synthetase of E. coli was covalently modified with the 3 ’terminal oxidized arginyl tRNA synthetase, and the ATP-PPi exchange activity and aminoacylation activity of the enzyme were completely lost. Analysis of the covalent complex of the enzyme with tRNA (?) By SDS-PAGE revealed that two distinctly different forms of ArgRS-tRNA (?) Complex were formed with the enzyme inactivation, which corresponded to two The mobility of different macromolecule band. This result is different from the results of other synthase affinity tags. ArgRS-tRNA (?) After RNase treatment, ATP-PPi exchange activity and aminoacylation activity were partially restored (less than 15%). Throughout the labeling process, the two vitality of the enzyme is closely related.