Bol'shakov M A, Ashikhmin A A, Makhneva Z K, Moskalenko A A
Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia.
Biochemistry (Mosc). 2015 Sep;80(9):1169-77. doi: 10.1134/S0006297915090072.
The effect of carotenoids on the assembly of LH2 complex in cells of the purple nonsulfur bacterium Rhodoblastus acidophilus was investigated. For this purpose, the bacterial culture was cultivated with an inhibitor of carotenoid biosynthesis - 71 µM diphenylamine (DPA). The inhibitor decreased the level of biosynthesis of the colored carotenoids in membranes by ~58%. It was found that a large amount of phytoene was accumulated in them. This carotenoid precursor was bound nonspecifically to LH2 complex and did not stabilize its structure. Thermostability testing of the isolated LH2 complex together with analysis of carotenoid composition revealed that the population of this complex was heterogeneous with respect to carotenoid composition. One fraction of the LH2 complex with carotenoid content around 90% remains stable and was not destroyed under heating for 15 min at 50°C. The other fraction of LH2 complex containing on average less than one molecule of carotenoid per complex was destroyed under heating, forming a zone of free pigments (and polypeptides). The data suggest that a certain part of the LH2 complexes is assembled without carotenoids in cells of the nonsulfur bacterium Rbl. acidophilus grown with DPA. These data contradict the fact that the LH2 complex from nonsulfur bacteria cannot be assembled without carotenoids, but on the other hand, they are in good agreement with the results demonstrated in our earlier studies of the sulfur bacteria Allochromatium minutissimum and Ectothiorhodospira haloalkaliphila. Carotenoidless LH2 complex was obtained from these bacteria with the use of DPA (Moskalenko, A. A., and Makhneva, Z. K. (2012) J. Photochem. Photobiol., 108, 1-7; Ashikhmin, A., et al. (2014) Photosynth. Res., 119, 291-303).
研究了类胡萝卜素对嗜酸红假单胞菌紫色非硫细菌细胞中LH2复合物组装的影响。为此,用类胡萝卜素生物合成抑制剂——71 μM二苯胺(DPA)培养细菌培养物。该抑制剂使膜中有色类胡萝卜素的生物合成水平降低了约58%。发现其中积累了大量的八氢番茄红素。这种类胡萝卜素前体与LH2复合物非特异性结合,并未稳定其结构。对分离的LH2复合物进行热稳定性测试并分析类胡萝卜素组成,结果表明该复合物群体在类胡萝卜素组成方面是异质的。LH2复合物的一部分类胡萝卜素含量约为90%,在50°C加热15分钟后仍保持稳定且未被破坏。LH2复合物的另一部分平均每个复合物含有的类胡萝卜素分子少于一个,在加热时被破坏,形成了游离色素(和多肽)区域。数据表明,在以DPA培养的非硫细菌嗜酸红假单胞菌细胞中,一定部分的LH2复合物是在没有类胡萝卜素的情况下组装的。这些数据与非硫细菌的LH2复合物在没有类胡萝卜素的情况下无法组装这一事实相矛盾,但另一方面,它们与我们早期对微小嗜色菌和嗜盐碱红螺菌的研究结果非常吻合。使用DPA从这些细菌中获得了无类胡萝卜素的LH2复合物(Moskalenko, A. A., and Makhneva, Z. K. (2012) J. Photochem. Photobiol., 108, 1 - 7; Ashikhmin, A., et al. (2014) Photosynth. Res., 119, 291 - 303)。