Utsugi J, Inaba K, Kuroda T, Tsuda M, Tsuchiya T
Department of Microbiology, Faculty of Pharmaceutical Sciences, Tsushima, Okayama 700, Japan.
Biochim Biophys Acta. 1998 Jul 9;1398(3):330-4. doi: 10.1016/s0167-4781(98)00058-x.
We cloned a gene for Na+/H+ antiporter from chromosomal DNA of Pseudomonas aeruginosa. Introduction of the gene into host Escherichia coli mutant cells lacking all of the major Na+/H+ antiporters enabled the cells to grow in the presence of 0.2 M NaCl, although the original host cells could not. Membrane vesicles prepared from cells of the transformant possessing the cloned gene showed Na+/H+ antiport activity. As a result of DNA sequencing, we found one open reading frame (nhaP). The deduced amino acid sequence suggests that the Na+/H+ antiporter (NhaP) of P. aeruginosa consists of 424 amino acid residues with molecular mass of 45486 Da, and hydropathy analysis suggested the presence of 12 putative transmembrane domains. We found no bacterial Na+/H+ antiporter which showed significant sequence similarity with the NhaP in the protein sequence database. The NhaP showed partial sequence similarity with animal Na+/H+ exchangers. Thus, the NhaP of P. aeruginosa is unique among bacterial antiporters.
我们从铜绿假单胞菌的染色体DNA中克隆了一个Na⁺/H⁺逆向转运蛋白基因。将该基因导入缺乏所有主要Na⁺/H⁺逆向转运蛋白的宿主大肠杆菌突变细胞后,这些细胞能够在0.2 M NaCl存在的情况下生长,而原始宿主细胞则不能。从含有克隆基因的转化体细胞制备的膜囊泡显示出Na⁺/H⁺逆向转运活性。通过DNA测序,我们发现了一个开放阅读框(nhaP)。推导的氨基酸序列表明,铜绿假单胞菌的Na⁺/H⁺逆向转运蛋白(NhaP)由424个氨基酸残基组成,分子量为45486 Da,亲水性分析表明存在12个推定的跨膜结构域。我们在蛋白质序列数据库中未发现与NhaP具有显著序列相似性的细菌Na⁺/H⁺逆向转运蛋白。NhaP与动物Na⁺/H⁺交换蛋白显示出部分序列相似性。因此,铜绿假单胞菌的NhaP在细菌逆向转运蛋白中是独特的。