Crielaard W, Driessen A J, Molenaar D, Hellingwerf K J, Konings W N
Department of Microbiology, University of Groningen, Haren, The Netherlands.
J Bacteriol. 1988 Apr;170(4):1820-4. doi: 10.1128/jb.170.4.1820-1824.1988.
Reaction centers of the phototrophic bacterium Rhodopseudomonas palustris were introduced as proton motive force-generating systems in membrane vesicles of two anaerobic bacteria. Liposomes containing reaction center-light-harvesting complex I pigment protein complexes were fused with membrane vesicles of Streptococcus cremoris or Clostridium acetobutylicum by freeze-thawing and sonication. Illumination of these fused membranes resulted in the generation of a proton motive force of approximately -110 mV. The magnitude of the proton motive force in these membranes could be varied by changing the light intensity. As a result of this proton motive force, amino acid transport into the fused membranes could be observed. The initial rate of leucine transport by membrane vesicles of S. cremoris increased exponentially with the proton motive force. An H+/leucine stoichiometry of 0.8 was determined from the steady-state level of leucine accumulation and the proton motive force, and this stoichiometry was found to be independent of the magnitude of the proton motive force. These results indicate that the introduction of bacterial reaction centers in membrane vesicles by the fusion procedure yields very attractive model systems for the study of proton motive force-consuming processes in membrane vesicles of (strict) anaerobic bacteria.
光合细菌沼泽红假单胞菌的反应中心作为质子动力产生系统被引入到两种厌氧细菌的膜囊泡中。通过冻融和超声处理,将含有反应中心 - 光捕获复合物I色素蛋白复合物的脂质体与嗜热链球菌或丙酮丁醇梭菌的膜囊泡融合。这些融合膜的光照导致产生约 -110 mV的质子动力。这些膜中质子动力的大小可以通过改变光强度来改变。由于这种质子动力,可以观察到氨基酸转运到融合膜中。嗜热链球菌膜囊泡转运亮氨酸的初始速率随质子动力呈指数增加。根据亮氨酸积累的稳态水平和质子动力确定H⁺/亮氨酸化学计量比为0.8,并且发现该化学计量比与质子动力的大小无关。这些结果表明,通过融合程序将细菌反应中心引入膜囊泡中,为研究(严格)厌氧细菌膜囊泡中质子动力消耗过程提供了非常有吸引力的模型系统。