Integration of fluorescence imaging with proteomics enables visualization and identification of metallo-proteomes in living cells. Issue 1 (10th November 2016)

• Record Type: Journal Article
• Authors: Lai, Yau-Tsz; Yang, Ya; Hu, Ligang; Cheng, Tianfan; Chang, Yuen-Yan; Koohi-Moghadam, Mohamad; Wang, Yuchuan; Xia, Jiang; Wang, Junwen; Li, Hongyan; Sun, Hongzhe
• Is Part Of: Metallomics. Volume 9:Issue 1(2017)
• Page Start: 38
• Page End: 47
• Publication Date: 2016-11-10
• Subjects: Metals -- Physiological effect -- Periodicals; 572.51
• DOI: 10.1039/c6mt00169f
• Languages: English
• ISSNs: 1756-5901

Abstract

Abstract : "Metal-tunable" fluorescent probe for tracking and editing metalloproteomes inside living cells. Abstract : Metalloproteins account for nearly one-third of proteins in proteomes. To date, the identification of metalloproteins relies mainly on protein purification and the subsequent characterization of bound metals, which often leads to losses of metal ions bound weakly and transiently. Herein, we developed a strategy to visualize and subsequently identify endogenous metalloproteins and metal-binding proteins in living cells via integration of fluorescence imaging with proteomics. We synthesized a "metal-tunable" fluorescent probe (denoted as M n + -TRACER) that rapidly enters cells to target proteins with 4–40 fold fluorescence enhancements. By using Ni 2+ -TRACER as an example, we demonstrate the feasibility of tracking Ni 2+ -binding proteins in vitro, while cellular small molecules exhibit negligible interference on the labeling. We identified 44 Ni 2+ -binding proteins from microbes using Helicobacter pylori as a showcase. We further applied Cu 2+ -TRACER to mammalian cells and found 54 Cu 2+ -binding proteins. The strategy we report here provides a great opportunity to track various endogenous metallo-proteomes and to mine potential targets of metallodrugs.