A conditional proteomics approach to identify proteins involved in zinc homeostasis
A conditional proteomics approach to identify proteins involved in zinc homeostasis
复制标题
DOI：
10.1038/nmeth.3998
复制
发表时间：
2016-09
期刊：
影响因子：
48
通讯作者：
Takayuki Miki;Masashi Awa;Yuki Nishikawa;Shigeki Kiyonaka;M. Wakabayashi;Y. Ishihama;I. Hamachi
中科院分区：
文献类型：
--
作者：
Takayuki Miki;Masashi Awa;Yuki Nishikawa;Shigeki Kiyonaka;M. Wakabayashi;Y. Ishihama;I. Hamachi
锌信号传导和动力学在许多生理反应和疾病中发挥着重要作用。为了详细了解锌的生理作用，在高浓度移动锌离子下全面鉴定蛋白质至关重要。我们开发了一种“条件蛋白质组学”方法，基于化学蛋白质组学策略来识别参与锌稳态的蛋白质，该策略利用设计的锌响应标记试剂来标记此类蛋白质，并通过定量质谱法进行鉴定。我们利用这种方法阐明了神经胶质瘤细胞中一氧化氮触发的氧化应激引起的锌稳态失调，并揭示了锌相关蛋白质组的动态变化。此外，我们将氧化应激产生的未知富锌囊泡表征为内质网和高尔基体相关囊泡。
Zinc signaling and dynamics play significant roles in many physiological responses and diseases. To understand the physiological roles of zinc in detail, comprehensive identification of proteins under high concentration of mobile zinc ion is crucial. We developed a 'conditional proteomics' approach to identify proteins involved in zinc homeostasis based on a chemical proteomic strategy that utilizes designer zinc-responsive labeling reagents to tag such proteins and quantitative mass spectrometry for their identification. We used this method to elucidate zinc dyshomeostasis induced by nitric-oxide-triggered oxidative stress in glioma cells, and we unveiled dynamic changes of the zinc-related proteomes. Moreover, we characterized unknown zinc-rich vesicles generated by oxidative stress as endoplasmic-reticulum- and Golgi-related vesicles.