Biogenesis of Extramitochondrial Iron-Sulfur Proteins in Eukaryotes

真核生物线粒体外铁硫蛋白的生物发生

• 批准号: 8965258
• 负责人: James Akira Wohlschlegel
• 金额: $ 24.54万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8965258
• 关键词: Amino Acids; Anemia; Apoproteins; Architecture; Binding; Biochemistry; Biogenesis; Cell Respiration; Cell physiology; Complex; Defect; Deuterium; Disease; Enzymes; Eukaryota; Golgi Apparatus; Hematological Disease; Hydrogen; Investigation; Iron; Iron-Sulfur Proteins; Knowledge; Laboratories; Link; Lipids; Maps; Mass Spectrum Analysis; Mediating; Metabolic Diseases; Molecular; Nature; Nuclear; Organelles; Pathogenesis; Pathway interactions; Play; Predisposition; Process; Protein S; Proteins; Proteomics; Recruitment Activity; Regulation; Research; Respiration; Role; Science; Site; Small Interfering RNA; Specificity; Staging; Sulfur; Syndrome; System; Testing; Therapeutic Intervention; Work; base; cancer genetics; cofactor; crosslink; insight; molecular recognition; nervous system disorder; novel; novel strategies; nucleic acid metabolism; nucleotide metabolism; protein complex; protein crosslink; research study; scaffold

Iron-sulfur (Fe-S) clusters are protein cofactors that play essential roles in virtually every facet of cellular physiology ranging from nucleic acid metabolism to aerobic respiration to the metabolism of nucleotides, amino acids, and lipids. The assembly and transfer of these cofactors to cytoplasmic and nuclear substrates is carried out by the cytoplasmic Fe-S protein assembly (CIA) machinery, a highly conserved biosynthetic pathway devoted to their biogenesis. Despite its central role in this process, our understanding of the CIA pathway remains lacking. My laboratory recently discovered a novel `CIA targeting complex' consisting of MMS19, FAM96B, and CIAO1 that functions at a late stage in the CIA pathway to recruit distinct sets of apoproteins to the CIA machinery and facilitate their assembly. In the proposed work, we will use a combination of biochemistry and proteomic mass spectrometry to investigate fundamental aspects of CIA targeting complex function. In specific aim 1, we will focus on understanding how the CIA targeting complex recognizes substrates. We will utilize both hydrogen/deuterium exchange mass spectrometry (HDX-MS) and cross-linking mass spectrometry (CX-MS) approaches to globally map protein interactions between the CIA targeting complex and its substrates in order to understand how molecular recognition and specificity are achieved. Specific aim 2 will examine the role of subcellular localization in the regulation of CIA targeting complex function. We find that the CIA targeting complex is localized to the ER-Golgi and now seek to understand the molecular basis, function, and significance of this localization. Investigation of these two aims will elucidate fundamental features regarding the regulation and function of the CIA targeting complex while potentially offering novels insights into how dysregulation of Fe-S cluster assembly contributes to a wide range of neurological, hematological, and metabolic disorders.

铁硫（Fe-S）簇是蛋白质辅助因子，在几乎所有的生物学过程中发挥重要作用。 从核酸代谢到有氧呼吸， 核苷酸、氨基酸和脂类的代谢。组装和转移 这些辅因子与细胞质和细胞核底物的结合是由细胞质 Fe-S蛋白组装（CIA）机制，一种高度保守的生物合成途径 致力于它们的生物起源尽管它在这一过程中发挥着核心作用，但我们对 中情局的途径仍然缺乏。我的实验室最近发现了一种新的“中情局” 由MMS 19、FAM 96 B和CIAO 1组成的“靶向复合物”，在晚期发挥作用。 阶段在CIA途径招募不同套脱辅基蛋白CIA机制， 方便他们的集会。在拟议的工作中，我们将使用生物化学的组合 和蛋白质组质谱法来研究CIA靶向的基本方面 复杂函数在具体目标1中，我们将重点了解中情局如何 靶向复合物识别底物。我们将利用氢/氘 交换质谱（HDX-MS）和交联质谱（CX-MS） 方法全局映射CIA靶向复合物之间的蛋白相互作用， 以了解分子识别和特异性是如何 办妥了一批具体目标2将检查亚细胞定位在调节中的作用， 中情局针对复杂功能我们发现中情局的目标群 ER-高尔基体，现在试图了解分子基础，功能和意义 这种本地化。对这两个目标的研究将阐明其基本特征 关于中央情报局目标复合体的监管和功能， 提供了关于Fe-S簇组装的失调如何有助于 广泛的神经系统、血液系统和代谢紊乱。