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首页> 美国卫生研究院文献>Plant Physiology >Properties of Substantially Chlorophyll-Free Pea Leaf Mitochondria Prepared by Sucrose Density Gradient Separation
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Properties of Substantially Chlorophyll-Free Pea Leaf Mitochondria Prepared by Sucrose Density Gradient Separation

机译：蔗糖密度梯度分离法制备的无叶绿素豌豆线粒体的性质

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摘要

Mitochondria isolated from pea leaves (Pisum sativum L. var Massey Gem) and purified on a linear sucrose density gradient were substantially free of contamination by Chl and peroxisomes. They showed high respiratory rates and good respiratory control and ADP/O ratios. Malate, glutamate, succinate, glycine, pyruvate, α-ketoglutarate, NADH, and NADPH were oxidized but little or no oxidation of citrate, isocitrate, or proline was detected. The oxidation of NADPH by the purified mitochondria did not occur via a transhydrogenase or phosphatase converting it to NADH. NADPH oxidation had an absolute requirement for added Ca2+, whereas NADH oxidation proceeded in its absence. In addition, oxidation of the two substrates showed different sensitivities to chelators and sulfhydryl reagents, and faster rates of O2 uptake were observed with both substrates than with either alone. This indicates that the NADPH dehydrogenase is distinct from the exogenous NADH dehydrogenase.
机译：从豌豆叶片（Pisum sativum L. var Massey Gem）分离并在线性蔗糖密度梯度上纯化的线粒体基本上不受Chl和过氧化物酶体的污染。他们表现出高呼吸频率和良好的呼吸控制以及ADP / O比值。苹果酸，谷氨酸，琥珀酸，甘氨酸，丙酮酸，α-酮戊二酸，NADH和NADPH被氧化，但柠檬酸，异柠檬酸或脯氨酸几乎没有氧化。纯化的线粒体不会通过转氢酶或磷酸酶将NADPH氧化为NADH而发生氧化。 NADPH氧化绝对需要添加Ca 2 + ，而NADH氧化则在不存在的情况下进行。此外，两种底物的氧化对螯合剂和巯基试剂的敏感性不同，与两种底物相比，两种底物的O2吸收速率更快。这表明NADPH脱氢酶不同于外源NADH脱氢酶。

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