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Identification of Cell-Surface Molecular Interactions under Living Conditions by Using the Enzyme-Mediated Activation of Radical Sources (EMARS) Method

by Koichi Honke 1,2,3,* and Norihiro Kotani 3,4
1
Department of Biochemistry, Kochi University Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan
2
Kochi System Glycobiology Center, Kochi University Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan
3
Center for Innovative and Translational Medicine, Kochi University Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan
4
Department of Biochemistry, Saitama Medical University, Iruma-gun, Saitama 350-0495, Japan
*
Author to whom correspondence should be addressed.
Sensors 2012, 12(12), 16037-16045; https://doi.org/10.3390/s121216037
Received: 8 October 2012 / Revised: 12 November 2012 / Accepted: 12 November 2012 / Published: 22 November 2012
(This article belongs to the Special Issue Live Cell-Based Sensors)
Important biological events associated with plasma membranes, such as signal transduction, cell adhesion, and protein trafficking, are mediated through the membrane microdomains. We have developed a novel method termed enzyme-mediated activation of radical sources (EMARS) to identify coclustering molecules on the cell surface under living conditions, which features a radical formation from an aryl azide reagent by horseradish peroxidase (HRP). For identification of molecules labeled by the EMARS reaction, antibody array system and mass spectrometry-based proteomics approaches are available. Spatio- temporally-regulated interaction between b1 integrin and ErbB4 involved in fibronectin-dependent cell migration and therapeutic antibody-stimulated interaction between FGFR3 and CD20 were discovered using the EMARS method. View Full-Text
Keywords: molecular interaction; membrane microdomains; lipid rafts; EMARS; HRP; receptor tyrosine kinases; antibody array; proteomics molecular interaction; membrane microdomains; lipid rafts; EMARS; HRP; receptor tyrosine kinases; antibody array; proteomics
MDPI and ACS Style

Honke, K.; Kotani, N. Identification of Cell-Surface Molecular Interactions under Living Conditions by Using the Enzyme-Mediated Activation of Radical Sources (EMARS) Method. Sensors 2012, 12, 16037-16045.

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