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

doi:10.3390/s121216037

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Sensors 2012, 12(12), 16037-16045; doi:10.3390/s121216037

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

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.

Received: 8 October 2012; in revised form: 12 November 2012 / Accepted: 12 November 2012 / Published: 22 November 2012

(This article belongs to the Special Issue Live Cell-Based Sensors)

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Abstract: 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.

Keywords: molecular interaction; membrane microdomains; lipid rafts; EMARS; HRP; receptor tyrosine kinases; antibody array; proteomics

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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.

AMA 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(12):16037-16045.

Chicago/Turabian Style

Honke, Koichi; Kotani, Norihiro. 2012. "Identification of Cell-Surface Molecular Interactions under Living Conditions by Using the Enzyme-Mediated Activation of Radical Sources (EMARS) Method." Sensors 12, no. 12: 16037-16045.

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