Shimamoto T, Inaba K, Thelen P, Ishikawa T, Goldberg E B, Tsuda M, Tsuchiya T
Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University.
J Biochem. 1994 Aug;116(2):285-90. doi: 10.1093/oxfordjournals.jbchem.a124521.
We isolated a mutant of Escherichia coli which was defective in an Na+/H+ antiporter and grew poorly under alkaline conditions [Ishikawa, T., Hama, H., Tsuda, T., and Tsuchiya, T. (1987) J. Biol. Chem. 262, 7443-7446]. Later, it was concluded that the defective Na+/H+ antiporter in the mutant was the NhaB system, and the nhaB gene was mapped to 25.6 min on the E. coli chromosome [Thelen, P., Tsuchiya, T., and Goldberg, E.B. (1991) J. Bacteriol. 173, 6553-6557]. We found that the NhaB-defective cells cannot grow in a high pH medium. Furthermore, intracellular pH in the mutant cells was almost the same as extracellular pH between 7.9 and 9.1, that is, intracellular pH was not regulated at this pH range. On the other hand, intracellular pH of the wild-type cells was maintained at about 7.6 when the extracellular pH was between 7.6 and 8.5. Thus, the NhaB Na+/H+ antiporter is essential for the regulation of intracellular pH under alkaline conditions in E. coli. Introduction of nhaA gene into the mutant cells increased Na+/H+ antiporter activity, but did not restore the defective growth and defective intracellular pH regulation under alkaline conditions.
我们分离出了一株大肠杆菌突变体,其钠离子/氢离子反向转运蛋白存在缺陷,在碱性条件下生长不良[石川,T.,滨,H.,津田,T.,和土屋,T.(1987年)《生物化学杂志》262卷,7443 - 7446页]。后来得出结论,该突变体中存在缺陷的钠离子/氢离子反向转运蛋白是NhaB系统,nhaB基因定位于大肠杆菌染色体上25.6分钟处[特伦,P.,土屋,T.,和戈德堡,E.B.(1991年)《细菌学杂志》173卷,6553 - 6557页]。我们发现,缺乏NhaB的细胞无法在高pH值培养基中生长。此外,在7.9至9.1之间,突变体细胞内的pH值与细胞外pH值几乎相同,也就是说,在此pH值范围内细胞内pH值不受调节。另一方面,当细胞外pH值在7.6至8.5之间时,野生型细胞的细胞内pH值维持在约7.6。因此,NhaB钠离子/氢离子反向转运蛋白对于大肠杆菌在碱性条件下调节细胞内pH值至关重要。将nhaA基因导入突变体细胞可增加钠离子/氢离子反向转运蛋白活性,但不能恢复碱性条件下生长缺陷和细胞内pH值调节缺陷。