Frost L S, Paranchych W, Willetts N S
J Bacteriol. 1984 Oct;160(1):395-401. doi: 10.1128/jb.160.1.395-401.1984.
The complete sequence of a 1.4-kilobase PstI fragment containing the F transfer genes traA, -L, and -E is presented. The traA reading frame has been located both genetically and by comparing the primary structure of F pilin (the traA product) predicted by the DNA sequence to the amino acid composition and sequence of N- and C-terminal peptides isolated from purified F pilin. Taken together, these data show that there is a leader peptide of 51 amino acids and that F pilin contains 70 amino acids, giving molecular weights of 13,200 for F propilin and 7,200 for mature F pilin. Secondary structure predictions for F pilin revealed a reverse turn that precedes the sequence Ala-Met-Ala51, a classic signal peptidase cleavage site. The N-terminal alanine residue is blocked by an acetyl group as determined by 1H-nuclear magnetic resonance spectroscopy. The traL and traE genes encode proteins of molecular weights 10,350 and 21,200, respectively. According to DNA sequence predictions, these proteins do not contain signal peptide leader sequences. Secondary structure predictions for these proteins are in accord with traLp and traEp being membrane proteins in which hydrophobic regions capable of spanning the membrane are linked by sequences that form turns and carry positively charged residues capable of interacting with the membrane surface.
本文给出了包含F转移基因traA、-L和-E的1.4千碱基对PstI片段的完整序列。通过遗传学方法以及将DNA序列预测的F菌毛蛋白(traA产物)一级结构与从纯化的F菌毛蛋白中分离出的N端和C端肽段的氨基酸组成及序列进行比较,确定了traA的阅读框。综合这些数据表明,存在一个由51个氨基酸组成的前导肽,F菌毛蛋白含有70个氨基酸,F前菌毛蛋白的分子量为13,200,成熟F菌毛蛋白的分子量为7,200。F菌毛蛋白的二级结构预测显示,在Ala-Met-Ala51序列之前有一个反向转角,这是一个典型的信号肽酶切割位点。通过1H-核磁共振光谱确定,N端丙氨酸残基被乙酰基封闭。traL和traE基因分别编码分子量为10,350和21,200的蛋白质。根据DNA序列预测,这些蛋白质不包含信号肽前导序列。这些蛋白质的二级结构预测表明,traLp和traEp是膜蛋白,其中能够跨膜的疏水区域通过形成转角并带有能够与膜表面相互作用的带正电荷残基的序列相连。