Maebashi Institute of Technology, Gunma 371-0816, Japan.
Talanta. 2011 Jul 15;85(1):425-9. doi: 10.1016/j.talanta.2011.04.003. Epub 2011 Apr 9.
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.
为了评估蛋白质-蛋白质相互作用，开发了一种新的伏安法，该方法使用标记有电活性化合物的蛋白质。伴刀豆球蛋白 A（ConA）是一种凝集素，可识别α-甘露糖残基。由于 ConA 要与具有高甘露糖糖链的卵清蛋白（OVA）结合，因此制备了标记有柔红霉素的 ConA 作为探针来监测结合。与 OVA 的结合是由于 ConA 的标签修饰引起的。结果，随着 OVA 浓度的增加，标记的 ConA 的电极响应降低。标记的 ConA 的电极响应在 1.5×10(-10)和 1.5×10(-9)M OVA 的范围内呈线性。1.5×10(-8)M 标记的 ConA 和 1.5×10(-10)M OVA 的相对标准偏差为 6.9％（n=5）。然后可以根据响应的变化方便地监测标记的 ConA-OVA 结合。相比之下，还研究了标记的 ConA 与没有特定糖链的蛋白质之间的相互作用。孵育几乎不会影响标记的 ConA 的峰电流。当向血清中添加几种浓度的 OVA 时，通过良好的回收率确定了这一点。因此，该方法可用于测量蛋白质-蛋白质相互作用。
