Krulwich T A, Guffanti A A, Bornstein R F, Hoffstein J
J Biol Chem. 1982 Feb 25;257(4):1885-9.
Activity of a Na+/H+ antiporter has been suggested to be critically involved in pH homeostasis in obligately alkalophilic bacteria (Krulwich, K. A., Mandel, K. G., Bornstein, R. F., and Guffanti, A. A. (1979) Biochem. Biophys. Res. Commun. 91, 58-62) and in Escherichia coli (Zilberstein, D., Padan, E., and Schuldiner, S. (1980) FEBS Lett. 116, 177-180). A concern with respect to these proposals has been the failure of either Bacillus alcalophilus or E. coli to exhibit a requirement for added Na+ for growth. Thus, it became of interest to examine Na+-coupled porter functions in obligately alkalophilic Bacillus firmus RAB, a species that exhibits an absolute requirement for added Na+ for growth at pH 10.5. In a comparative study using membrane vesicles from B. alcalophilus and B. firmus RAB it was found that both the Na+/H+ antiporter and the Na+/alpha-aminoisobutyric acid symporter from the "Na+-requiring" species had much lower apparent affinities for Na+ than corresponding porters from B. alcalophilus. At high concentrations of Na+, the porters from the two species were functionally similar. These findings support the argument that the absence of a growth requirement for added Na+ may reflect an ability of at least some bacteria to effectively utilize and recycle the available levels of Na+ that contaminate all media, rather than reflect true Na+ independence. Studies with a nonalkalophilic derivative of B. firmus RAB confirmed earlier findings with B. alcalophilus of a pleiotropic loss of Na+ coupling to porters in nonalkalophilic mutants.
据推测,Na⁺/H⁺逆向转运蛋白的活性在专性嗜碱细菌(Krulwich, K. A., Mandel, K. G., Bornstein, R. F., and Guffanti, A. A. (1979) Biochem. Biophys. Res. Commun. 91, 58 - 62)和大肠杆菌(Zilberstein, D., Padan, E., and Schuldiner, S. (1980) FEBS Lett. 116, 177 - 180)的pH稳态中起着关键作用。关于这些推测的一个问题是,嗜碱芽孢杆菌或大肠杆菌在生长过程中都没有表现出对添加Na⁺的需求。因此,研究专性嗜碱的坚强芽孢杆菌RAB中与Na⁺偶联的转运蛋白功能变得很有意义,该菌株在pH 10.5时生长绝对需要添加Na⁺。在一项使用嗜碱芽孢杆菌和坚强芽孢杆菌RAB膜囊泡的比较研究中发现,“需要Na⁺”物种的Na⁺/H⁺逆向转运蛋白和Na⁺/α-氨基异丁酸同向转运蛋白对Na⁺的表观亲和力远低于嗜碱芽孢杆菌相应的转运蛋白。在高浓度Na⁺下,这两个物种的转运蛋白在功能上相似。这些发现支持了这样一种观点,即对添加Na⁺没有生长需求可能反映了至少一些细菌能够有效利用和循环污染所有培养基的可用Na⁺水平,而不是反映真正的Na⁺独立性。对坚强芽孢杆菌RAB的非嗜碱衍生物的研究证实了早期对嗜碱芽孢杆菌的研究结果,即在非嗜碱突变体中,Na⁺与转运蛋白的偶联出现多效性丧失。