Department of Molecular Biology, School of Pharmacy, Iwate Medical University, Nishitokuta, Yahaba, Shiwa-gun, Iwate, Japan.
J Bacteriol. 2013 Nov;195(21):4873-8. doi: 10.1128/JB.00542-13. Epub 2013 Aug 23.
The c subunit of Streptococcus mutans ATP synthase (FoF1) is functionally exchangeable with that of Escherichia coli, since E. coli with a hybrid FoF1 is able to grow on minimum succinate medium through oxidative phosphorylation. E. coli F1 bound to the hybrid Fo with the S. mutans c subunit showed N,N'-dicyclohexylcarbodiimide-sensitive ATPase activity similar to that of E. coli FoF1. Thus, the S. mutans c subunit assembled into a functional Fo together with the E. coli a and b subunits, forming a normal F1 binding site. Although the H(+) pathway should be functional, as was suggested by the growth on minimum succinate medium, ATP-driven H(+) transport could not be detected with inverted membrane vesicles in vitro. This observation is partly explained by the presence of an acidic residue (Glu-20) in the first transmembrane helix of the S. mutans c subunit, since the site-directed mutant carrying Gln-20 partly recovered the ATP-driven H(+) transport. Since S. mutans is recognized to be a primary etiological agent of human dental caries and is one cause of bacterial endocarditis, our system that expresses hybrid Fo with the S. mutans c subunit would be helpful to find antibiotics and chemicals specifically directed to S. mutans.
变形链球菌 ATP 合酶的 c 亚基(FoF1)与大肠杆菌的 c 亚基在功能上可互换,因为具有杂种 FoF1 的大肠杆菌能够通过氧化磷酸化在最小琥珀酸盐培养基上生长。与 S. mutans c 亚基结合的 E. coli F1 与杂种 Fo 结合显示出类似于 E. coli FoF1 的 N,N'-二环己基碳化二亚胺敏感的 ATP 酶活性。因此,S. mutans c 亚基与大肠杆菌的 a 和 b 亚基一起组装成功能性 Fo,形成正常的 F1 结合位点。尽管 H(+)途径应该是功能性的,正如在最小琥珀酸盐培养基上生长所表明的那样,但在体外使用翻转膜囊泡不能检测到 ATP 驱动的 H(+)转运。这一观察结果部分解释了 S. mutans c 亚基第一跨膜螺旋中存在酸性残基(Glu-20),因为携带 Gln-20 的定点突变体部分恢复了 ATP 驱动的 H(+)转运。由于变形链球菌被认为是人类龋齿的主要病原体之一,也是细菌性心内膜炎的原因之一,因此我们表达具有 S. mutans c 亚基的杂种 Fo 的系统将有助于找到专门针对 S. mutans 的抗生素和化学物质。