Morstadt Lucia, Gräber Peter, De Pascalis Luca, Kleinig Hans, Speth Volker, Beyer Peter
Zentrum für Angewandte Biowissenschaften, Universität Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
Planta. 2002 May;215(1):134-40. doi: 10.1007/s00425-001-0724-y. Epub 2002 Feb 19.
Mature chromoplasts from daffodil (Narcissus pseudonarcissus) flowers, although devoid of thylakoid structures, contain immunologically detectable alpha-subunits of ATP-synthase (H(+)-transporting ATP phosphohydrolase; EC 3.6.3.14). To show the presence of the entire functional protein complex, chromoplast membrane proteins were solubilized and reconstituted in phosphatidylcholine liposomes. The membranes were energized by an acid-base transition in the presence of a K(+)/valinomycin diffusion potential, and the initial rate of ATP synthesis was measured with a luciferin/luciferase assay. In addition, by demonstrating NADPH-dependent ATP synthesis, we show that an NAD(P)H-dependent respiratory redox pathway in chromoplasts, previously identified as an important constituent of the carotene desaturation system, proceeds concomitant with membrane energization.
来自水仙（Narcissus pseudonarcissus）花朵的成熟质体，虽然没有类囊体结构，但含有免疫可检测的ATP合酶α亚基（H⁺转运ATP磷酸水解酶；EC 3.6.3.14）。为了证明整个功能蛋白复合物的存在，将质体膜蛋白溶解并在磷脂酰胆碱脂质体中重构。在存在K⁺/缬氨霉素扩散电位的情况下，通过酸碱转变使膜获得能量，并使用荧光素/荧光素酶测定法测量ATP合成的初始速率。此外，通过证明NADPH依赖性ATP合成，我们表明质体中先前被鉴定为胡萝卜素去饱和系统重要组成部分的NAD(P)H依赖性呼吸氧化还原途径与膜能量化同时进行。
