Abstract
Membrane proteomics, the large-scale global analysis of membrane proteins, is often constrained by the efficiency of separating and extracting membrane proteins. Recent approaches involve conjugating membrane proteins with the small molecule biotin and using the receptor streptavidin to extract the labelled proteins. Despite the many advantages of this method, several shortcomings remain, including potential contamination by endogenously biotinylated molecules and interference by streptavidin during analytical stages. Here, we report a supramolecular fishing method for membrane proteins using the synthetic receptor–ligand pair cucurbit[7]uril–1-trimethylammoniomethylferrocene (CB[7]–AFc). CB[7]-conjugated beads selectively capture AFc-labelled proteins from heterogeneous protein mixtures, and AFc-labelling of cells results in the efficient capture of membrane proteins by these beads. The captured proteins can be recovered easily at room temperature by treatment with a strong competitor such as 1,1′-bis(trimethylammoniomethyl)ferrocene. This synthetic but biocompatible host–guest system may be a useful alternative to streptavidin–biotin for membrane proteomics as well as other biological and biotechnological applications.
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Acknowledgements
This work was supported by the Creative Research Initiative and Brain Korea 21 Program of the Korean Ministry of Education, Science, and Technology (MOEST), the World Class University (WCU) program through the Korea Science and Engineering Foundation funded by MOEST (project no. R31-2008-000-10059-0). The authors also thank J. Mark Kim for helpful discussions.
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K.K. and S.H.R conceived and designed the project. D.W.L., K.M.P., B.M., K.S., P.S. and N.S. performed the synthesis. D.W.L. and H.J. performed the protein and cell experiments. D.W.L., K.M.P. and S.H.H. analysed the data. T.L. performed mass spectrometry. J.K. provided invaluable advice. K.K., D.W.L. and K.M.P. wrote the paper.
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Lee, DW., Park, K., Banerjee, M. et al. Supramolecular fishing for plasma membrane proteins using an ultrastable synthetic host–guest binding pair. Nature Chem 3, 154–159 (2011). https://doi.org/10.1038/nchem.928
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DOI: https://doi.org/10.1038/nchem.928
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