ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, Crawley, WA, 6009, Australia.
Plant Mol Biol. 2010 Feb;72(3):331-42. doi: 10.1007/s11103-009-9573-z. Epub 2009 Nov 19.
Complex II plays a central role in mitochondrial metabolism as a component of both the electron transport chain and the tricarboxylic acid cycle. However, the composition and function of the plant enzyme has been elusive and differs from the well-characterised enzymes in mammals and bacteria. Herewith, we demonstrate that mitochondrial Complex II from Arabidopsis and rice differ significantly in several aspects: (1) Stability-Rice complex II in contrast to Arabidopsis is not stable when resolved by native electrophoresis and activity staining. (2) Composition-Arabidopsis complex II contains 8 subunits, only 7 of which have homologs in the rice genome. SDH 1 and 2 subunits display high levels of amino acid identity between two species, while the remainder of the subunits are not well conserved at a sequence level, indicating significant divergence. (3) Gene expression-the pairs of orthologous SDH1 and SDH2 subunits were universally expressed in both Arabidopsis and rice. The very divergent genes for SDH3 and SDH4 were co-expressed in both species, consistent with their functional co-ordination to form the membrane anchor. The plant-specific SDH5, 6 and 7 subunits with unknown functions appeared to be differentially expressed in both species. (4) Biochemical regulation -succinate-dependent O(2) consumption and SDH activity of isolated Arabidopsis mitochondria were substantially stimulated by ATP, but a much more minor effect of ATP was observed for the rice enzyme. The ATP activation of succinate-dependent reduction of DCPIP in frozen-thawed and digitonin-solubilised mitochondrial samples, and with or without the uncoupler CCCP, indicate that the differential ATP effect on SDH is not via the protonmotive force but likely due to an allosteric effect on the plant SDH enzyme itself, in contrast to the enzyme in other organisms.
复合物 II 作为电子传递链和三羧酸循环的组成部分,在线粒体代谢中起着核心作用。然而,植物酶的组成和功能一直难以捉摸,与哺乳动物和细菌中特征良好的酶不同。在此,我们证明来自拟南芥和水稻的线粒体复合物 II 在几个方面存在显著差异:(1)稳定性 - 与拟南芥不同,水稻复合物 II 在通过天然电泳和活性染色解析时不稳定。(2)组成 - 拟南芥复合物 II 包含 8 个亚基,其中只有 7 个在水稻基因组中有同源物。SDH1 和 2 亚基在两个物种之间显示出高度的氨基酸同一性,而其余的亚基在序列水平上没有很好的保守性,表明存在显著的差异。(3)基因表达 - 两个物种中普遍表达了一对同源的 SDH1 和 SDH2 亚基。SDH3 和 SDH4 的非常不同的基因在两个物种中共同表达,这与它们形成膜锚的功能协调一致。具有未知功能的植物特异性 SDH5、6 和 7 亚基似乎在两个物种中表现出差异表达。(4)生化调节 - 分离的拟南芥线粒体中的琥珀酸盐依赖性 O2 消耗和 SDH 活性被 ATP 显著刺激,但在水稻酶中观察到 ATP 的影响要小得多。在冷冻-解冻和去垢剂溶解的线粒体样品中,以及在有无解偶联剂 CCCP 的情况下,ATP 依赖性琥珀酸盐依赖性 DCPIP 的还原表明,SDH 的 ATP 激活不是通过质子动力,而是可能由于对植物 SDH 酶本身的变构效应,与其他生物体中的酶不同。