Cancer-Related NEET Proteins Transfer 2Fe-2S Clusters to Anamorsin, a Protein Required for Cytosolic Iron-Sulfur Cluster Biogenesis.
Cancer-Related NEET Proteins Transfer 2Fe-2S Clusters to Anamorsin, a Protein Required for Cytosolic Iron-Sulfur Cluster Biogenesis.
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DOI：
10.1371/journal.pone.0139699
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Jennings PA
中科院分区：
文献类型：
--
作者：
Lipper CH;Paddock ML;Onuchic JN;Mittler R;Nechushtai R;Jennings PA
铁硫簇生物合成是由不同的蛋白质组装系统执行的。哺乳动物有两个系统，线粒体Fe-S簇组装系统（ISC）和胞质组装系统（CIA），它们通过未知的机制连接。2 Fe-2S蛋白NEET家族的人类成员营养剥夺自噬因子-1（NAF-1）和mitoNEET（mNT）位于线粒体和细胞质之间的界面。这些蛋白质与癌细胞增殖有关，它们可以将其2Fe-2S簇转移到标准的apo受体蛋白上。在这里，我们报告了两种NEET蛋白的第一个生理2Fe-2S簇受体，即人Anamorsin（也称为细胞因子诱导的凋亡抑制剂-1; CIAPIN-1）。Anamorsin是一种电子传递蛋白，含有两个铁硫簇结合位点，是细胞溶质Fe-S簇组装所必需的。我们表明，使用UV-Vis光谱，NAF-1和mNT都可以将其2Fe-2S簇转移到apo-Anamorsin，其二级速率常数与其他已知的人2Fe-2S转移蛋白相似。使用生物层干涉测量法检测NEET蛋白与脱辅基Anamorsin的直接蛋白质-蛋白质相互作用。此外，电喷雾质谱的簇转移制备的holo-Anamorsin表明，它接受了它的两个2Fe-2S团簇从NEX。我们建议，mNT和NAF-1可以提供连接线粒体ISC系统和CIA的平行路线。在线粒体中组装的2Fe-2S簇被NEET蛋白接收，并在需要时转移到Anamorsin，激活CIA。
Iron-sulfur cluster biogenesis is executed by distinct protein assembly systems. Mammals have two systems, the mitochondrial Fe-S cluster assembly system (ISC) and the cytosolic assembly system (CIA), that are connected by an unknown mechanism. The human members of the NEET family of 2Fe-2S proteins, nutrient-deprivation autophagy factor-1 (NAF-1) and mitoNEET (mNT), are located at the interface between the mitochondria and the cytosol. These proteins have been implicated in cancer cell proliferation, and they can transfer their 2Fe-2S clusters to a standard apo-acceptor protein. Here we report the first physiological 2Fe-2S cluster acceptor for both NEET proteins as human Anamorsin (also known as cytokine induced apoptosis inhibitor-1; CIAPIN-1). Anamorsin is an electron transfer protein containing two iron-sulfur cluster-binding sites that is required for cytosolic Fe-S cluster assembly. We show, using UV-Vis spectroscopy, that both NAF-1 and mNT can transfer their 2Fe-2S clusters to apo-Anamorsin with second order rate constants similar to those of other known human 2Fe-2S transfer proteins. A direct protein-protein interaction of the NEET proteins with apo-Anamorsin was detected using biolayer interferometry. Furthermore, electrospray mass spectrometry of holo-Anamorsin prepared by cluster transfer shows that it receives both of its 2Fe-2S clusters from the NEETs. We propose that mNT and NAF-1 can provide parallel routes connecting the mitochondrial ISC system and the CIA. 2Fe-2S clusters assembled in the mitochondria are received by NEET proteins and when needed transferred to Anamorsin, activating the CIA.