Moor N A, Ankilova V N, Favre A, Lavrik O I
Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia.
Biochemistry (Mosc). 1998 Sep;63(9):1051-6.
A photoreactive tRNAPhe derivative containing a 4-thiouridine residue at the 3'-end (tRNAPhe-s4U-75) was prepared by tRNA nucleotidyltransferase-mediated incorporation of s4UMP into a tRNAPhe transcript lacking the 3'-terminal dinucleotide. The resulting tRNAPhe-s4U-75 was covalently bound to phenylalanyl-tRNA synthetase from Thermus thermophilus, and all criteria of an affinity modification were met. The main products of modification displaying various electrophoretic mobilities were formed by binding tRNAPhe-s4U-75 to the beta-subunit (major) of the enzyme. These data suggest that the nucleotide found at position 75 of tRNAPhe interacts with the beta-subunit of phenylalanyl-tRNA synthetase.
通过tRNA核苷酸转移酶介导将s4UMP掺入缺乏3'-末端二核苷酸的tRNAPhe转录本中，制备了一种在3'-末端含有4-硫尿苷残基的光反应性tRNAPhe衍生物（tRNAPhe-s4U-75）。所得的tRNAPhe-s4U-75与嗜热栖热菌的苯丙氨酰-tRNA合成酶共价结合，满足了亲和修饰的所有标准。修饰的主要产物呈现出各种电泳迁移率，是通过将tRNAPhe-s4U-75与该酶的β亚基（主要）结合形成的。这些数据表明，tRNAPhe第75位的核苷酸与苯丙氨酰-tRNA合成酶的β亚基相互作用。
