title: Electrochemically monitoring the binding of concanavalin A and ovalbumin.
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.
authors.affiliation: Maebashi Institute of Technology, Gunma 371-0816, Japan. kzsuga@maebashi-it.ac.jp
authors.forename: Kazuharu
authors.initials: K
authors.lastname: Sugawara
authors.forename: A
authors.initials: A
authors.lastname: Yugami
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Kadoya
authors.forename: Kohei
authors.initials: K
authors.lastname: Hosaka
mesh_terms: D003208:Concanavalin A / Q000737:chemistry*; D003630:Daunorubicin; D055664:Electrochemical Techniques / Q000379:methods*; D015335:Molecular Probes; D010047:Ovalbumin / Q000097:blood / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D015335:Molecular Probes; D003208:Concanavalin A; D010047:Ovalbumin; D003630:Daunorubicin