Integrative Metallomic Approach to Identify Metalloproteins

Abstract

It is crucial to identify metal-protein interactions at a proteome-wide scale (1) which are difficult due to certain weak and transient interactions. We developed an integrated approach consisting of gel electrophoresis and inductively coupled plasma mass spectrometry, LA-ICP-MS, IMAC, fluorescence and bioinformatics to identify metal-associated proteins using bismuth antiulcer drug as an example (2,3). We have identified metal-associated proteins as well as to quantify the metals for fast metallome/proteome-wide profiling of metal-binding proteins. Metal-tunable fluorescence probe can track both His-tagged proteins and metal-binding proteins in live cells (4). We also revealed selective toxicity of bismuth(III) drugs towards certain pathogens but not humans. Over 90% of bismuth was found to be passively absorbed, conjugated to glutathione and transported into vesicles by MRP transporter (5). We further established a bioinformatic method which allows potential metal-binding proteins both sequentially and spaciously to be searched. The versatile modes of action provide a rational basis in understanding the high efficacy and low antibiotic resistance of this class of bismuth drugs (6). We also show that an integrative metallomic approach represents a powerful tool for metals in biology and medicine as well as pharmacology of metallodrugs. We thank the Research Grants Council of Hong Kong, Innovative Technology Fund, NSFC and the University of Hong Kong for financial support. References [1] H. Sun (ed.) Biological chemistry of arsenic, antimony and bismuth 2011, John Wiley. [2] L.G. Hu, T.F. Cheng, G.B. Jiang, H. Sun etc, Angew Chem Int Ed 2013, 52, 4916. [3] H. Li, H. Sun, Curr Opin Chem Biol. 2012, 16, 74. [4] Y.T. Lai, Y.Y. Chang, L. Hu, Y. Yang, A. Chao, H. Sun etc, PNAS, 2015, 112, 2948. [5] Y. Hong, Y.T. Lai, G.C. Chan, H. Sun, PNAS 2015, 112, 3211. [6] Y.C., Wang, C.N.Tsang, F. Xu, J.W. Wang, , I.K. Chu, H. Li, H. Sun, Chem Comm 2015, 51, 16479.