Electrochemically monitoring the binding of concanavalin A and ovalbumin |
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Authors: | Sugawara Kazuharu Yugami A Kadoya Toshihiko Hosaka Kohei |
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Affiliation: | a Maebashi Institute of Technology, Gunma 371-0816, Japan b Faculty of Education, Gunma University, Gunma 371-8510, Japan c Basic Sciences for Medicine, Gunma University School of Health Sciences, Gunma 371-8511, Japan |
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Abstract: | To evaluate protein-protein interactions, a new voltammetric method was developed using a protein labeled with an electroactive compound. Concanavalin A (ConA), which is a lectin, recognizes α-mannose residues. Because the ConA was to be bound to ovalbumin (OVA), which has a high-mannose sugar chain, ConA labeled with daunomycin was prepared as the probe to monitor the binding. The binding to OVA was caused by the label modification of the ConA. As a result, the electrode response of the labeled ConA decreased as the OVA concentration increased. The electrode response of the labeled ConA was linearly over the range of 1.5 × 10−10 and 1.5 × 10−9 M OVA. The relative standard deviation of 1.5 × 10−8 M labeled ConA and 1.5 × 10−10 M OVA was 6.9% (n = 5). The labeled ConA-OVA binding could then be conveniently monitored based on the change in response. In contrast, interactions between the labeled ConA and a protein with no specific sugar chain also were investigated. Incubation scarcely influenced the peak current of the labeled ConA. When several concentrations of OVA were added to a serum, good recovery determined it. Consequently, this method could be applied to the measurement of protein-protein interactions. |
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Keywords: | Ovalbumin Concanavalin A Protein-protein interaction Voltammetry |
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