Hall C V, vanCleemput M, Muench K H, Yanofsky C
J Biol Chem. 1982 Jun 10;257(11):6132-6.
The complete nucleotide sequence of trpS, the structural gene for Escherichia coli tryptophanyl-tRNA synthetase, was determined using a plasmid carrying the structural gene. From the single open reading frame of correct length and orientation we deduced an amino acid sequence consistent with the amino acid composition of the purified protein. In addition, previously sequenced peptides representing 52% of the protein were readily aligned with regions of the deduced sequence. The deduced amino acid sequence of the E. coli enzyme is 60% homologous with the sequence of the enzyme from Bacillus stearothermophilus. Using currently available procedures we predicted the secondary structure for the enzyme from each organism and compared these structures to those of the two aminoacyl-tRNA synthetases whose three-dimensional structures have been determined. We used a convenient plasmid recombination procedure to map the regional locations of missense mutations within trpS that have characteristic effects on the properties of the enzyme.
利用携带大肠杆菌色氨酰 - tRNA合成酶结构基因的质粒,测定了trpS(该酶的结构基因)的完整核苷酸序列。从具有正确长度和方向的单一开放阅读框中,我们推导了与纯化蛋白的氨基酸组成一致的氨基酸序列。此外,先前测序的、占该蛋白52%的肽段很容易与推导序列的区域对齐。大肠杆菌该酶的推导氨基酸序列与嗜热脂肪芽孢杆菌的酶序列有60%的同源性。我们使用现有的方法预测了每种生物该酶的二级结构,并将这些结构与已确定三维结构的两种氨酰 - tRNA合成酶的结构进行了比较。我们采用一种简便的质粒重组程序来定位trpS内错义突变的区域位置,这些突变对酶的性质有特征性影响。