Kreutzer R, Kern D, Giegé R, Rudinger J
Lehrstuhl für Biochemie, Universität Bayreuth, Germany.
Nucleic Acids Res. 1995 Nov 25;23(22):4598-602. doi: 10.1093/nar/23.22.4598.
The phosphates of the tRNA(Phe) transcript from Thermus thermophilus interacting with the cognate synthetase were determined by footprinting. Backbone bond protection against cleavage by iodine of the phosphorothioate-containing transcripts was found in the anticodon stem-loop, the D stem-loop and the acceptor stem and weak protection was also seen in the variable loop. Most of the protected phosphates correspond to regions around known identity elements of tRNA(Phe). Enhancement of cleavage at certain positions indicates bending of tRNAPhe upon binding to the enzyme. When applied to the three-dimensional model of tRNA(Phe) from yeast the majority of the protections occur on the D loop side of the molecule, revealing that phenylalanyl-tRNA synthetase has a rather complex and novel pattern of interaction with tRNAPhe, differing from that of other known class II aminoacyl-tRNA synthetases.
通过足迹法确定了嗜热栖热菌的tRNA（Phe）转录本与同源合成酶相互作用的磷酸盐。在反密码子茎环、D茎环和受体茎中发现了对含硫代磷酸酯转录本碘切割的主链键保护，在可变环中也观察到了弱保护。大多数受保护的磷酸盐对应于tRNA（Phe）已知识别元件周围的区域。某些位置切割增强表明tRNAPhe与酶结合时发生弯曲。当应用于酵母tRNA（Phe）的三维模型时，大多数保护发生在分子的D环一侧，表明苯丙氨酰-tRNA合成酶与tRNAPhe具有相当复杂和新颖的相互作用模式，与其他已知的II类氨酰-tRNA合成酶不同。
