Kossmann M, Wolff C, Manson M D
Department of Biology, Texas A&M University, College Station 77843.
J Bacteriol. 1988 Oct;170(10):4516-21. doi: 10.1128/jb.170.10.4516-4521.1988.
The maltose chemoreceptor in Escherichia coli consists of the periplasmic maltose-binding protein (MBP) and the Tar signal transducer, which is localized in the cytoplasmic membrane. We previously isolated strains containing malE mutations that cause specific defects in the chemotactic function of MBP. Four of these mutations have now been characterized by DNA sequence analysis. Two of them replace threonine at residue 53 of MBP with isoleucine (MBP-TI53), one replaces an aspartate at residue 55 with asparagine (MBP-DN55), and the fourth replaces threonine at residue 345 with isoleucine (MBP-TI345). The chemotactic defects of MBP-TI53 and MBP-DN55, but not of MBP-TI345, are suppressed by mutations in the tar gene. Of the tar mutations, the most effective suppressor (isolated independently three times) replaces Arg-73 of Tar with tryptophan. Two other tar mutations that disrupt the aspartate chemoreceptor function of Tar also suppress the maltose taxis defects associated with MBP-TI53 and MBP-DN55. One of these mutations introduces glutamine at residue 73 of Tar, the other replaces arginine at residue 69 of Tar with cysteine. These results suggest that regions of MBP that include residues 53 to 55 and residue 345 are important for the interaction with Tar. In turn, arginines at residues 69 and 73 of Tar must be involved in the recognition of maltose-bound MBP and/or in the production of the attractant signal generated by Tar in response to maltose-bound MBP.
大肠杆菌中的麦芽糖化学感受器由周质麦芽糖结合蛋白(MBP)和位于细胞质膜中的Tar信号转导器组成。我们之前分离出了含有malE突变的菌株,这些突变在MBP的趋化功能中导致特定缺陷。现在通过DNA序列分析对其中四个突变进行了表征。其中两个突变将MBP第53位残基的苏氨酸替换为异亮氨酸(MBP-TI53),一个将第55位残基的天冬氨酸替换为天冬酰胺(MBP-DN55),第四个将第345位残基的苏氨酸替换为异亮氨酸(MBP-TI345)。MBP-TI53和MBP-DN55的趋化缺陷(而非MBP-TI345的缺陷)被tar基因中的突变所抑制。在tar突变中,最有效的抑制子(独立分离出三次)将Tar的Arg-73替换为色氨酸。另外两个破坏Tar天冬氨酸化学感受器功能的tar突变也抑制了与MBP-TI53和MBP-DN55相关的麦芽糖趋化缺陷。其中一个突变在Tar的第73位残基引入谷氨酰胺,另一个将Tar第69位残基的精氨酸替换为半胱氨酸。这些结果表明,MBP中包括第53至55位残基和第345位残基的区域对于与Tar的相互作用很重要。反过来,Tar第69位和第73位残基的精氨酸必定参与对结合麦芽糖的MBP的识别和/或参与Tar响应结合麦芽糖的MBP产生的吸引信号。