Functional nucleic acid and Nucleotide sugar
Comprehensively labeling of cell surface molecular clustering in living cells
EMARS (Enzyme-Mediated Activation of Radical Sources) is a labeling method that labels molecules comprehensively within a limited distance (300nm) from the probed molecules on which HRP is set. The EMARS method proposes to be a powerful tool to elucidate molecular clustering on the cell surface of living cells in various contexts.
EMARS reagent and EMARS reaction
First step: setting HRP on cell membrane
Second step: EMARS reaction (a case of setting HRP by antibody)
Schematic diagram of the in vivo EMARS method Living cells are treated with HRP-conjugated cognitive molecules, and subsequently treated with EMARS reagent which consist of aryl azide compound bonded with labeling compound. Aryl azide the part of EMARS reagent is converted to active radical species by HRP and attack the molecules located within a limited distance from HRP set on a given molecule on the cell surface and label the coclustered molecules. After the EMARS reaction, the membrane proteins are solubilized and the labeled molecules by EMARS reagents are identified by using ordinary method of protein analysis such as antibody array.
Last step: analysis of FITC labeled molecule
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Application protocols in preparation
 Jiang,S., Kotani,N., Honke,K., et.al., A proteomics approach to the cell-surface interactome using the enzyme-mediated activation of radical sources reaction. Proteomics 2012,12,54-62
 Taniguchi,N.,Commentary-Searching for partners, Proteomics 2012,12,9-10
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