Pogoryelov Denys, Reichen Christian, Klyszejko Adriana L, Brunisholz René, Muller Daniel J, Dimroth Peter, Meier Thomas
Max-Planck-Institute of Biophysics, Max-von-Laue-Strasse 3, 60438 Frankfurt am Main, Germany.
J Bacteriol. 2007 Aug;189(16):5895-902. doi: 10.1128/JB.00581-07. Epub 2007 Jun 1.
We isolated the c rings of F-ATP synthases from eight cyanobacterial strains belonging to four different taxonomic classes (Chroococcales, Nostocales, Oscillatoriales, and Gloeobacteria). These c rings showed different mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), probably reflecting their molecular masses. This supposition was validated with the previously characterized c(11), c(14), and c(15) rings, which migrated on SDS-PAGE in proportion to their molecular masses. Hence, the masses of the cyanobacterial c rings can conveniently be deduced from their electrophoretic mobilities and, together with the masses of the c monomers, allow the calculation of the c ring stoichiometries. The method is a simple and fast way to determine stoichiometries of SDS-stable c rings and hence a convenient means to unambiguously determine the ion-to-ATP ratio, a parameter reflecting the bioenergetic efficacy of F-ATP synthases. AFM imaging was used to prove the accuracy of the method and confirmed that the c ring of Synechococcus elongatus SAG 89.79 is a tridecameric oligomer. Despite the high conservation of the c-subunit sequences from cyanobacterial strains from various environmental groups, the stoichiometries of their c rings varied between c(13) and c(15). This systematic study of the c-ring stoichiometries suggests that variability of c-ring sizes might represent an adaptation of the individual cyanobacterial species to their particular environmental and physiological conditions. Furthermore, the two new examples of c(15) rings underline once more that an F(1)/F(o) symmetry mismatch is not an obligatory feature of all F-ATP synthases.
我们从属于四个不同分类类群(色球藻目、念珠藻目、颤藻目和粘球藻纲)的八个蓝藻菌株中分离出了F-ATP合酶的c环。这些c环在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)上显示出不同的迁移率,这可能反映了它们的分子量。这一推测通过先前已表征的c(11)、c(14)和c(15)环得到了验证,它们在SDS-PAGE上的迁移率与其分子量成正比。因此,蓝藻c环的质量可以方便地从其电泳迁移率中推导出来,并且与c单体的质量一起,可以计算c环的化学计量比。该方法是确定SDS稳定的c环化学计量比的一种简单快速的方法,因此是明确确定离子与ATP比率的一种便捷手段,该比率是反映F-ATP合酶生物能量效率的一个参数。原子力显微镜成像用于证明该方法的准确性,并证实了细长聚球藻SAG 89.79的c环是一种十三聚体寡聚体。尽管来自不同环境群体的蓝藻菌株的c亚基序列具有高度保守性,但其c环的化学计量比在c(13)和c(15)之间有所不同。对c环化学计量比的这项系统研究表明,c环大小的变异性可能代表了各个蓝藻物种对其特定环境和生理条件的一种适应。此外,c(15)环的两个新例子再次强调,F(1)/F(o)对称性不匹配并非所有F-ATP合酶的必然特征。