Stehling Oliver, Smith Paul M, Biederbick Annette, Balk Janneke, Lill Roland, Mühlenhoff Ulrich
Institut für Zytobiologie und Zytopathologie, Philipps-Universität Marburg, Germany.
Methods Mol Biol. 2007;372:325-42. doi: 10.1007/978-1-59745-365-3_24.
Iron-sulfur (Fe-S) clusters are cofactors of many proteins that are involved in central biochemical pathways, such as oxidative phosphorylation, photosynthesis, and amino acid biosynthesis. The assembly of these cofactors and the maturation of Fe-S proteins require complex cellular machineries in all kingdoms of life. In eukaryotes, Fe-S protein biogenesis is an essential process, and mitochondria perform a primary role in synthesis. Defects in Fe-S protein maturation in yeast result in respiratory deficiency and auxotrophies for certain amino acids and vitamins that require Fe-S proteins for their biosynthesis. Frequently, heme biosynthesis is also affected. The present compendium describes assays for the analysis of de novo Fe-S cluster and heme formation, cellular iron homeostasis, and the activity of Fe-S cluster- and heme-containing enzymes. These approaches are crucial to elucidate the mechanisms underlying the maturation of Fe-S proteins and may aid in the identification of new members of this evolutionary ancient process.
铁硫（Fe-S）簇是许多参与中心生化途径的蛋白质的辅因子，这些途径包括氧化磷酸化、光合作用和氨基酸生物合成。在所有生命王国中，这些辅因子的组装以及Fe-S蛋白的成熟都需要复杂的细胞机制。在真核生物中，Fe-S蛋白生物合成是一个必不可少的过程，线粒体在合成中起主要作用。酵母中Fe-S蛋白成熟的缺陷会导致呼吸缺陷以及对某些氨基酸和维生素的营养缺陷型，这些氨基酸和维生素的生物合成需要Fe-S蛋白。通常，血红素生物合成也会受到影响。本综述描述了用于分析从头合成Fe-S簇和血红素形成、细胞铁稳态以及含Fe-S簇和血红素的酶活性的测定方法。这些方法对于阐明Fe-S蛋白成熟的潜在机制至关重要，并且可能有助于鉴定这一古老进化过程的新成员。
