Boston College, Department of Chemistry, Chestnut Hill, MA, USA.
Methods Mol Biol. 2024;2839:261-289. doi: 10.1007/978-1-0716-4043-2_16.
Iron-sulfur (Fe-S) clusters are essential redox-active metallocofactors participating in electron transfer, radical chemistry, primary metabolism, and gene regulation. Successful trafficking and incorporation of Fe-S clusters into target proteins are critical to proper cellular function. While biophysical studies of isolated Fe-S proteins provide insight into the structure and function of these inorganic cofactors, few strategies currently exist to directly interrogate Fe-S cluster binding within a cellular environment. Here, we describe a chemoproteomic platform to report on Fe-S cluster incorporation and occupancy directly within a native proteome, enabling the characterization of Fe-S biogenesis pathways and the identification of undiscovered Fe-S proteins.
铁硫 (Fe-S) 簇是参与电子传递、自由基化学、初级代谢和基因调控的重要氧化还原活性金属辅因子。Fe-S 簇成功转运和掺入靶蛋白对于细胞的正常功能至关重要。虽然对分离的 Fe-S 蛋白的生物物理研究提供了对这些无机辅因子结构和功能的深入了解，但目前几乎没有直接在细胞环境中检测 Fe-S 簇结合的策略。在这里，我们描述了一种化学生物组学平台，可以直接在天然蛋白质组中报告 Fe-S 簇的掺入和占据情况，从而能够对 Fe-S 生物发生途径进行表征并鉴定未发现的 Fe-S 蛋白。
