Inoue-Kashino Natsuko, Takahashi Takeshi, Ban Akiko, Sugiura Miwa, Takahashi Yuichiro, Satoh Kazuhiko, Kashino Yasuhiro
Department of Life Science, University of Hyogo, Ako-gun, Hyogo 678-1297, Japan.
Photosynth Res. 2008 Oct-Dec;98(1-3):323-35. doi: 10.1007/s11120-008-9340-z. Epub 2008 Aug 8.
Ycf12 (Psb30) is a small hydrophobic subunit of photosystem II (PS II) complexes found in the cyanobacterium, Thermosynechococcus elongatus. However, earlier intense proteomic analysis on the PS II complexes from the cyanobacterium, Synechocystis 6803, could not detect Psb30. In this work, we generated a mutant of Synechocystis 6803 in which a hexa-histidine tag was fused to the C-terminus of Synechocystis Psb30. The mutant accumulated fully functional PS II complexes. Purification of Psb30 by metal affinity chromatography from thylakoid extracts resulted in co-purification of an oxygen-evolving PS II complex with normal subunit composition. This result indicates that Psb30 is expressed and stably associated with the PS II complex in Synechocystis. The histidine-tagged Psb30 in the purified PS II complex was not detected by staining or anti-polyhistidine antibodies. We also generated a mutant in which ycf12 was disrupted. The mutant grew photosynthetically and showed no significant phenotype under moderate growth conditions. Purified PS II complexes from the disruptant showed an oxygen-evolving activity comparable to wild type under low irradiance. However, it showed a remarkably lower activity than wild type under high irradiance. Thus Psb30 is required for the efficient function of PS II complexes, particularly under high irradiance conditions.
Ycf12(Psb30)是在嗜热栖热放线菌中发现的光系统II(PS II)复合物的一个小的疏水性亚基。然而,早期对集胞藻6803的PS II复合物进行的深入蛋白质组学分析未能检测到Psb30。在这项工作中,我们构建了集胞藻6803的一个突变体,其中六组氨酸标签融合到集胞藻Psb30的C末端。该突变体积累了功能完全正常的PS II复合物。通过金属亲和色谱从类囊体提取物中纯化Psb30,导致具有正常亚基组成的放氧PS II复合物共纯化。这一结果表明Psb30在集胞藻中表达并与PS II复合物稳定结合。纯化的PS II复合物中带组氨酸标签的Psb30通过染色或抗多组氨酸抗体未被检测到。我们还构建了一个ycf12被破坏的突变体。该突变体在中等生长条件下进行光合作用生长,且未表现出明显的表型。从该破坏突变体中纯化的PS II复合物在低光照下显示出与野生型相当的放氧活性。然而,在高光照下,它的活性比野生型显著降低。因此,Psb30是PS II复合物高效功能所必需的,特别是在高光照条件下。