Analysis of the pathogenic human mitochondrial mutation ND1/3460, and mutations of strictly conserved residues in its vicinity, using the bacterium Paracoccus denitrificans.
Analysis of the pathogenic human mitochondrial mutation ND1/3460, and mutations of strictly conserved residues in its vicinity, using the bacterium Paracoccus denitrificans.
复制标题
使用脱氮副球菌分析致病性人类线粒体突变 ND1/3460 及其附近严格保守残基的突变。
DOI：
10.1021/bi9810555
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发表时间：
1998
期刊：
影响因子：
2.9
通讯作者：
M. Finel
中科院分区：
文献类型：
--
作者：
V. Zickermann;B. Barquera;M. Wikström;M. Finel
人类线粒体ND1/3460突变将复合体I的ND1亚单位中的Ala52改变为Thr，并导致Leber遗传性视神经病变(LHON)[Huoponen等人。(1991)上午J.嗯。吉内。48,1147]。我们使用了脱氮副球藻复合体I的细菌对应物NDH-1，来研究ND1亚基突变对酶活性的影响。LHON突变以及其附近几个严格保守的氨基酸突变被引入NDH-1的NQO8亚基中，NDH-1是ND1的细菌同源物。对突变株在六矿金(鱼藤酮不敏感)和泛醌-1(鱼藤酮敏感)存在下的酶活性进行了测定。此外，还研究了所选择的突变酶与泛醌-1、泛醌-2和去环路联醌相互作用的动力学。结果表明，突变残基在络合物I还原泛醌过程中起着重要作用。
The human mitochondrial ND1/3460 mutation changes Ala52 to Thr in the ND1 subunit of Complex I, and causes Leber's hereditary optic neuropathy (LHON) [Huoponen et al. (1991) Am. J. Hum. Genet. 48, 1147]. We have used a bacterial counterpart of Complex I, NDH-1 from Paracoccus denitrificans, for studying the effect of mutations in the ND1 subunit on the enzymatic activity. The LHON mutation as well as several other mutations in strictly conserved amino acids in its vicinity were introduced into the NQO8 subunit of NDH-1, a bacterial homologue of ND1. The enzymatic activity of the mutants in the presence of hexammineruthenium (rotenone-insensitive) and ubiquinone-1 (rotenone-sensitive) were assayed. In addition, the kinetics of the interaction of selected mutant enzymes with ubiquinone-1, ubiquinone-2, and decylubiquinone was studied. The results suggest that the mutated residues play an important role in ubiquinone reduction by Complex I.