Purification and characterization of NADH dehydrogenase complex from Paracoccus denitrificans.
Purification and characterization of NADH dehydrogenase complex from Paracoccus denitrificans.
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DOI：
10.1016/0003-9861(86)90731-9
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发表时间：
1986-11
影响因子：
3.9
通讯作者：
Takao Yagi
中科院分区：
文献类型：
--
作者：
Takao Yagi
用NaBr抽提和NAD-琼脂糖柱纯化，从嗜氧副球菌细胞质膜中分离到一种NADH脱氢酶复合物。分离的NADH脱氢酶复合物的NADH-泛醌-1还原酶活性比NaBr提取物高约10倍。该制剂由10种（6种主要和4种次要）不同的多肽组成，并且缺乏氧化磷酸化系统的其他酶复合物的可识别组分和活性特征。纯化的酶含有非共价结合的FMN，非血红素铁，和酸不稳定的硫化物。FMN、非血红素铁和酸不稳定硫化物的比例为1：13~14：11~12，表明存在多个铁硫簇合物。分离的NADH脱氢酶复合物与牛心脏线粒体复合物I和来自其的蛋白质组分的抗血清发生交叉反应，表明在Paracoccus酶中存在与完整的复合物I及其铁硫蛋白和可能的疏水蛋白组分中的抗原位点相似的抗原位点。
An NADH dehydrogenase complex was isolated from the plasma membranes of aerobically grownParacoccus denitrificanscells by extraction with NaBr and purification on an NAD-agarose column. The NADH-ubiquinone-1 reductase activity of the isolated NADH dehydrogenase complex was about 10 times higher than that of the NaBr extract. The preparation was composed of 10 (6 major and 4 minor) unlike polypeptides, and lacked identifiable components and activities characteristic of other enzyme complexes of the oxidative phosphorylation system. The purified enzyme contained noncovalently bound FMN, nonheme iron, and acid-labile sulfide. The ratio of FMN to nonheme iron to acid-labile sulfide was 1:13~14:11~12, suggestive of the presence of multiple iron-sulfur clusters. The isolated NADH dehydrogenase complex cross-reacted with antisera to beef heart mitochondrial complex I and protein fraction derived therefrom, indicating the presence in theParacoccusenzyme of antigenic sites similar to those in the intact complex I and its iron-sulfur protein and possibly hydrophobic protein fractions.