Mass spectrometry of iron-sulfur cluster regulatory proteins

铁硫簇调节蛋白的质谱分析

• 批准号: 1801873
• 金额: --
• 依托单位: University of East Anglia
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1801873
• 关键词: Mass; spectrometry; iron; sulfur; cluster

Iron-sulfur (FeS) cluster-containing regulatory proteins play key roles in sensing and responding to environmental change. The cluster functions as the sensory module, undergoing reaction that leads to protein conformational changes that enable the signal to be transduced. The [4Fe-4S] cluster containing WhiB-like (Wbl) family of regulators (named after the first discovered WhiB protein) are found in the actinobacteria, a phylum of Gram-positive bacteria that includes Streptomyces and important pathogens such as Mycobacterium tuberculosis. These proteins play key roles in cell development, controlling sporulation in Streptomyces and the transition into dormancy in M. tuberculosis. Recent work suggests that Wbl proteins may function through sensing nitric oxide (NO) and this project is focussed on understanding this reaction.The principal technique that will be used is native mass spectrometry (MS), which detects intact proteins in their folded states where non-covalent interactions (including protein-metallocofactor) are preserved upon vaporisation/ionisation. Thus, native MS has the potential to observe not only resting states of enzymes/proteins, but also intermediates in which substrates are non-covalently attached. Initial experiments indicate that native MS can provide an unprecedented level of detail about the reactions of iron-sulfur clusters with physiologically relevant small molecule reactants such as nitric oxide.

含有铁硫 (FeS) 簇的调节蛋白在感知和响应环境变化方面发挥着关键作用。该簇起到感觉模块的作用，发生反应导致蛋白质构象变化，从而使信号得以转导。包含 WhiB 样 (Wbl) 调节因子家族（以第一个发现的 WhiB 蛋白命名）的 [4Fe-4S] 簇存在于放线菌中，放线菌是革兰氏阳性菌门，包括链霉菌和结核分枝杆菌等重要病原体。这些蛋白质在细胞发育、控制链霉菌孢子形成和结核分枝杆菌进入休眠状态的转变中发挥关键作用。最近的研究表明，Wbl 蛋白可能通过感测一氧化氮 (NO) 发挥作用，该项目的重点是了解这种反应。将使用的主要技术是天然质谱 (MS)，它检测处于折叠状态的完整蛋白质，其中在汽化/电离时保留非共价相互作用（包括蛋白质-金属辅因子）。因此，天然 MS 不仅可以观察酶/蛋白质的静息状态，还可以观察底物非共价连接的中间体。初步实验表明，天然 MS 可以提供有关铁硫簇与生理相关小分子反应物（例如一氧化氮）反应的前所未有的详细信息。