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首页> 外文期刊>Biochemistry >The Human Iron-Sulfur Assembly Complex Catalyzes the Synthesis of [2Fe-2S] Clusters on ISCU2 That Can Be Transferred to Acceptor Molecules
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The Human Iron-Sulfur Assembly Complex Catalyzes the Synthesis of [2Fe-2S] Clusters on ISCU2 That Can Be Transferred to Acceptor Molecules

机译：人铁 - 硫组合络合物催化在ISCU2上的[2FE-2S]簇的合成，其可转移到受体分子

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摘要

Ironsulfur (FeS) clusters are essential protein cofactors for most life forms. In human mitochondria, the core FeS biosynthetic enzymatic complex (called SDUF) consists of NFS1, ISD11, ISCU2, and frataxin (FXN) protein components. Few mechanistic details about how this complex synthesizes FeS clusters and how these clusters are delivered to targets are known. Here circular dichroism and Mossbauer spectroscopies were used to reveal details of the FeS cluster assembly reaction on the SDUF complex. SDUF reactions generated [2Fe-2S] cluster intermediates that readily converted to stable [2Fe-2S] clusters bound to uncomplexed ISCU2. Similar reactions that included the apo FeS acceptor protein human ferredoxin (FDX1) resulted in formation of [2Fe-2S]-ISCU2 rather than [2Fe-2S]-FDX1. Subsequent addition of dithiothreitol (DTT) induced transfer of the cluster from ISCU2 to FDX1, suggesting that [2Fe-2S]-ISCU2 is an intermediate. Reactions that initially included DTT rapidly generated [2Fe-2S]-FDX1 and bypassed formation of [2Fe-2S]-ISCU2. In the absence of apo-FDX1, incubation of [2Fe-2S]-ISCU2 with DTT generated [4Fe-4S]-ISCU2 species. Together, these results conflict with a recent report of stable [4Fe-4S] cluster formation on the SDUF complex. Rather, they support a model in which SDUF builds transient [2Fe-2S] cluster intermediates that generate clusters on sulfur-containing molecules, including uncomplexed ISCU2. Additional small molecule or protein factors are required for the transfer of these clusters to FeS acceptor proteins or the synthesis of [4Fe-4S] clusters.
机译：Ironsulfur（FES）簇是大多数寿命形式的必需蛋白质辅助因子。在人体线粒体中，核心FES生物合成酶复合物（称为SDUF）由NFS1，ISD11，ISCU2和Frataxin（FXN）蛋白质组分组成。关于这种复杂的细节很少有关如何合成FES群集以及如何将这些集群传递给目标。在这里，使用圆形二中间和母蛋白光谱揭示SDUF复合物对FES簇组装反应的细节。 SDUF反应产生[2FE-2S]簇中间体，其容易转化为与未复制的ISCU2结合的稳定的[2FE-2S]簇。类似的反应包括APO FES受体蛋白人的福兰蛋白（FDX1）导致形成[2FE-2S] -iscu2而不是[2FE-2S] -FDX1。随后加入二硫醇（DTT）诱导簇从ISCU2转移到FDX1，表明[2FE-2S] -iscu2是中间体。最初包括DTT的反应快速产生[2FE-2S] -Fdx1并绕过[2FE-2S] -iscu2的形成。在不存在APO-FDX1的情况下，用DTT产生[2FE-2S] -iscu2的温育[4FE-4S] -iscu2种。在一起，这些结果与最近关于SDUF复合体稳定[4FE-4S]群集形成的报告发生冲突。相反，它们支持其中SDUF构建瞬态[2FE-2S]群体中间体，该模型在含硫分子上产生簇，包括未复合的ISCU2。需要额外的小分子或蛋白质因子来转移这些簇以FES受体蛋白或[4FE-4S]簇的合成。

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