Expression and Functional Roles of the Two Distinct NDH-1 Complexes and the Carbon Acquisition Complex NdhD3/NdhF3/CupA/Sll1735 in Synechocystis sp PCC 6803
Expression and Functional Roles of the Two Distinct NDH-1 Complexes and the Carbon Acquisition Complex NdhD3/NdhF3/CupA/Sll1735 in Synechocystis sp PCC 6803
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DOI：
10.1105/tpc.104.026526
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发表时间：
2004-12
期刊：
影响因子：
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通讯作者：
Pengpeng Zhang;N. Battchikova;T. Jansén;Jens Appel;T. Ogawa;E. Aro
中科院分区：
文献类型：
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作者：
Pengpeng Zhang;N. Battchikova;T. Jansén;Jens Appel;T. Ogawa;E. Aro
为了研究多功能NAD（P）H脱氢酶1型（NDH-1）复合物的（共）表达、相互作用和膜定位及其在碳获取、循环光系统I和呼吸中的作用，我们在不同CO2和pH条件下培养了集胞藻6803野生型和特异性ndh基因敲除突变体，然后对其膜蛋白复合物进行了蛋白质组学分析。典型的NDH-1复合物以NDH-1 L（大）和NDH-1 M（中等大小）为代表，位于类囊体膜上。Δ NdhD 1/D2突变体中缺失的NDH-1 L复合物是光合异养生长的先决条件，因此显然参与细胞呼吸。在低CO2条件下生长的Δ NdhD 1/D2突变体中，在黑暗中NDH-1 M的量和P700+再还原的速率与野生型相似，而在NDH-1 L和NDH-1 M均缺失的M55突变体（ΔNdhB）中，P700+再还原的速率非常缓慢。NDH-1 S（小）复合物，定位于类囊体膜，仅由NdhD 3，NdhF 3，CupA和Sll 1735组成，在野生型以及Δ NdhD 1/D2和M55中在低CO2下被强烈诱导。与野生型和Δ NdhD 1/D2（表现出正常的CO2吸收）相比，即使存在NDH-1 S复合物，M55也无法吸收CO2。相反，Δ NdhD 3/D4突变体也不能吸收CO2，缺乏NDH-1 S，但在低CO2下表现出野生型水平的NDH-1 M。这些结果表明，NDH-1 S和NDH-1 M都是CO2吸收所必需的，NDH-1 M是一种功能性复合物。我们还表明，Na+/HCO 3 −转运蛋白（SbtA复合物）位于质膜上，在野生型和突变体中在低CO2下被强烈诱导。
To investigate the (co)expression, interaction, and membrane location of multifunctional NAD(P)H dehydrogenase type 1 (NDH-1) complexes and their involvement in carbon acquisition, cyclic photosystem I, and respiration, we grew the wild type and specific ndh gene knockout mutants of Synechocystis sp PCC 6803 under different CO2 and pH conditions, followed by a proteome analysis of their membrane protein complexes. Typical NDH-1 complexes were represented by NDH-1L (large) and NDH-1M (medium size), located in the thylakoid membrane. The NDH-1L complex, missing from the ΔNdhD1/D2 mutant, was a prerequisite for photoheterotrophic growth and thus apparently involved in cellular respiration. The amount of NDH-1M and the rate of P700+ rereduction in darkness in the ΔNdhD1/D2 mutant grown at low CO2 were similar to those in the wild type, whereas in the M55 mutant (ΔNdhB), lacking both NDH-1L and NDH-1M, the rate of P700+ rereduction was very slow. The NDH-1S (small) complex, localized to the thylakoid membrane and composed of only NdhD3, NdhF3, CupA, and Sll1735, was strongly induced at low CO2 in the wild type as well as in ΔNdhD1/D2 and M55. In contrast with the wild type and ΔNdhD1/D2, which show normal CO2 uptake, M55 is unable to take up CO2 even when the NDH-1S complex is present. Conversely, the ΔNdhD3/D4 mutant, also unable to take up CO2, lacked NDH-1S but exhibited wild-type levels of NDH-1M at low CO2. These results demonstrate that both NDH-1S and NDH-1M are essential for CO2 uptake and that NDH-1M is a functional complex. We also show that the Na+/HCO3− transporter (SbtA complex) is located in the plasma membrane and is strongly induced in the wild type and mutants at low CO2.