Peng Lianwei, Shimizu Hideyuki, Shikanai Toshiharu
Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
J Biol Chem. 2008 Dec 12;283(50):34873-9. doi: 10.1074/jbc.M803207200. Epub 2008 Oct 14.
The chloroplast NAD(P)H dehydrogenase (NDH) complex is involved in photosystem I (PSI) cyclic and chlororespiratory electron transport in higher plants. Although biochemical and genetic evidence for its subunit composition has accumulated, it is not enough to explain the complexes putative activity of NAD(P)H-dependent plastoquinone reduction. We analyzed the NDH complex by using blue native PAGE and found that it interacts with PSI to form a novel supercomplex. Mutants lacking NdhL and NdhM accumulated a pigment-protein complex with a slightly lower molecular mass than that of the NDH-PSI supercomplex; this may be an intermediate supercomplex including PSI. This intermediate is unstable in mutants lacking NdhB, NdhD, or NdhF, implying that it includes some NDH subunits. Analysis of thylakoid membrane complexes using sucrose density gradient centrifugation supported the presence of the NDH-PSI supercomplex in vivo. Although the NDH complex exists as a monomer in etioplasts, it interacts with PSI to form a supercomplex within 48 h during chloroplast development.
叶绿体NAD(P)H脱氢酶(NDH)复合体参与高等植物光系统I(PSI)的循环以及叶绿体呼吸电子传递。尽管关于其亚基组成的生化和遗传学证据已不断积累,但仍不足以解释该复合体假定的依赖NAD(P)H的质体醌还原活性。我们通过蓝色非变性聚丙烯酰胺凝胶电泳(blue native PAGE)分析了NDH复合体,发现它与PSI相互作用形成一种新型超复合体。缺乏NdhL和NdhM的突变体积累了一种色素蛋白复合体,其分子量略低于NDH-PSI超复合体;这可能是一个包含PSI的中间超复合体。这种中间体在缺乏NdhB、NdhD或NdhF的突变体中不稳定,这意味着它包含一些NDH亚基。使用蔗糖密度梯度离心法对类囊体膜复合体进行分析,证实了体内存在NDH-PSI超复合体。尽管NDH复合体在黄化质体中以单体形式存在,但在叶绿体发育过程中,它会在48小时内与PSI相互作用形成超复合体。