Localization and functional characterization of the alternative oxidase in Naegleria.

Localization and functional characterization of the alternative oxidase in Naegleria.
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DOI:
10.1111/jeu.12908
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发表时间:
2022-07
期刊:
The Journal of eukaryotic microbiology
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其他
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交替氧化酶(AOX)是一种蛋白质,在典型(细胞色素介导的)呼吸链被抑制的情况下参与支持克雷布斯循环的酶促反应,同时允许维持细胞生长和生存所需的代谢过程。在真核生物中,交替氧化酶具有分散的分布,并且发现于植物、真菌和原生生物中,包括耐格里属物种。耐格里属物种是自由生活的单细胞阿米巴鞭毛虫,包括福氏耐格里属的致病物种,即所谓的“食脑阿米巴”。使用多学科的方法,我们的目的是了解AOX的进化,定位和功能以及在Naegleria生物学中的作用。我们的分析表明AOX存在于该属的最后一个共同祖先中,结构预测表明所有功能残基也存在于耐格里属物种中。使用细胞和生物化学技术,我们还功能性地描述了N. gruberi在其线粒体中的AOX,我们证明了它的失活会影响它的增殖。因此,我们讨论了该蛋白质在耐格里属物种中存在的益处,沿着其在福氏耐格里属中的潜在致病作用。我们预测,我们的研究结果将引领新的探索,以了解耐格里属和其他自由生活的亲属的细胞生物学,代谢和进化。
The alternative oxidase (AOX) is a protein involved in supporting enzymatic reactions of the Krebs cycle in instances when the canonical (cytochrome‐mediated) respiratory chain has been inhibited, while allowing for the maintenance of cell growth and necessary metabolic processes for survival. Among eukaryotes, alternative oxidases have dispersed distribution and are found in plants, fungi, and protists, including Naegleria ssp. Naegleria species are free‐living unicellular amoeboflagellates and include the pathogenic species of N. fowleri, the so‐called “brain‐eating amoeba.” Using a multidisciplinary approach, we aimed to understand the evolution, localization, and function of AOX and the role that plays in Naegleria's biology. Our analyses suggest that AOX was present in last common ancestor of the genus and structure prediction showed that all functional residues are also present in Naegleria species. Using cellular and biochemical techniques, we also functionally characterize N. gruberi's AOX in its mitochondria, and we demonstrate that its inactivation affects its proliferation. Consequently, we discuss the benefits of the presence of this protein in Naegleria species, along with its potential pathogenicity role in N. fowleri. We predict that our findings will spearhead new explorations to understand the cell biology, metabolism, and evolution of Naegleria and other free‐living relatives.
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