Cytosolic iron-sulfur protein assembly system identifies clients by a C-terminal tripeptide.
Cytosolic iron-sulfur protein assembly system identifies clients by a C-terminal tripeptide.
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胞浆铁硫蛋白组装系统通过 C 端三肽识别客户。
DOI：
10.1101/2023.05.19.541488
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Pierik,AntonioJ
中科院分区：
文献类型：
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作者：
Marquez,MelissaD;Greth,Carina;Buzuk,Anastasiya;Liu,Yaxi;Blinn,CatharinaM;Beller,Simone;Leiskau,Laura;Hushka,Anthony;Wu,Kassandra;Nur,Kübra;Netz,DailiJ;Perlstein,DeborahL;Pierik,AntonioJ
真核细胞质Fe-S蛋白组装（CIA）机制将铁-硫（Fe-S）簇插入细胞质和核蛋白中。在最后的成熟步骤中，Fe-S簇通过CIA靶向复合物（CTC）转移到脱辅基蛋白。然而，客户蛋白的分子识别决定簇是未知的。我们发现，保守的[LIM]-[DES]-[WF]-COO-三肽存在于超过四分之一的客户端或其适配器的C-末端。当存在时，这种靶向复合物识别（TCR）基序对于体外结合CTC和体内指导Fe-S簇递送是必要的和足够的。值得注意的是，这种TCR信号的融合使得能够通过募集CIA机制在非天然蛋白质上工程化簇成熟。我们的研究推进了我们对Fe-S蛋白成熟的理解，并为含Fe-S酶的生物工程新途径铺平了道路。
The eukaryotic cytosolic Fe–S protein assembly (CIA) machinery inserts iron–sulfur (Fe–S) clusters into cytosolic and nuclear proteins. In the final maturation step, the Fe–S cluster is transferred to the apo-proteins by the CIA-targeting complex (CTC). However, the molecular recognition determinants of client proteins are unknown. We show that a conserved [LIM]-[DES]-[WF]-COO–tripeptide is present at the C-terminus of more than a quarter of clients or their adaptors. When present, this targeting complex recognition (TCR) motif is necessary and sufficient for binding to the CTC in vitro and for directing Fe–S cluster delivery in vivo. Remarkably, fusion of this TCR signal enables engineering of cluster maturation on a nonnative protein via recruitment of the CIA machinery. Our study advances our understanding of Fe–S protein maturation and paves the way for bioengineering novel pathways containing Fe–S enzymes.