An antibody toolbox to track complex I assembly defines AIF's mitochondrial function.
An antibody toolbox to track complex I assembly defines AIF's mitochondrial function.
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DOI：
10.1083/jcb.202001071
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发表时间：
2020-10-05
期刊：
影响因子：
--
通讯作者：
Owusu-Ansah E
中科院分区：
文献类型：
--
作者：
Murari A;Rhooms SK;Goparaju NS;Villanueva M;Owusu-Ansah E
在线粒体复合体I(CI)组装过程中，独立形成的组装中间体(AIs)相互融合形成成熟的复合体。Murari等人。生成一个包含21种不同线粒体蛋白抗体的工具箱，并使用经典的果蝇遗传学和人工智能的免疫印迹来定义人工智能在脑梗塞生物发生中的作用。全面跟踪果蝇复杂的I(CI)生物发生的组装中间产物(AI)的能力将使人们能够描述各种CI调节因子调控CI组装的精确机制(S)。因此，我们产生了21种针对不同线粒体蛋白的新抗体，并利用这一资源通过跟踪当AIF表达受损时观察到的AI谱来表征AIF调节CI生物发生的机制。我们发现，当AIF-Mia40易位复合体被破坏时，CI中将电子转移到泛醌的部分被合成，但无法在CI生物合成途径中进行。这与Mia40底物MIC19在线粒体内积累的减少有关。重要的是，MIC19或MIC60，线粒体接触位点和脊组织系统(MICOS)的组成部分，完全概括了当AIF被抑制时观察到的AI谱。因此，AIF对CI组装的影响主要是由于MICOS复合体在线粒体内的转运受到损害。
During mitochondrial complex I (CI) assembly, independently formed assembly intermediates (AIs) merge with each other to form the mature complex. Murari et al. generate a toolbox of 21 antibodies to various mitochondrial proteins and use classical Drosophila genetics and immunoblotting of AIs to define AIF’s role in CI biogenesis. An ability to comprehensively track the assembly intermediates (AIs) of complex I (CI) biogenesis in Drosophila will enable the characterization of the precise mechanism(s) by which various CI regulators modulate CI assembly. Accordingly, we generated 21 novel antibodies to various mitochondrial proteins and used this resource to characterize the mechanism by which apoptosis-inducing factor (AIF) regulates CI biogenesis by tracking the AI profile observed when AIF expression is impaired. We find that when the AIF–Mia40 translocation complex is disrupted, the part of CI that transfers electrons to ubiquinone is synthesized but fails to progress in the CI biosynthetic pathway. This is associated with a reduction in intramitochondrial accumulation of the Mia40 substrate, MIC19. Importantly, knockdown of either MIC19 or MIC60, components of the mitochondrial contact site and cristae organizing system (MICOS), fully recapitulates the AI profile observed when AIF is inhibited. Thus, AIF’s effect on CI assembly is principally due to compromised intramitochondrial transport of the MICOS complex.
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影响因子：
64.8
作者：
Fiedorczuk, Karol;Letts, James A.;Degliesposti, Gianluca;Kaszuba, Karol;Skehel, Mark;Sazanov, Leonid A.
通讯作者：
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影响因子：
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作者：
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作者：
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DOI：
10.1016/j.bbamcr.2007.12.005
发表时间：
2008-04-01
影响因子：
5.1
作者：
Hell, Kai
通讯作者：
Hell, Kai
影响因子：
29
作者：
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通讯作者：
Nijtmans, Leo