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  • 09월 04일 17시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제114회 대한화학회 학술발표회, 총회 및 기기전시회 안내 A synthetic chemosensor based electrochemical assay of protein kinase on a microchip

2014년 8월 28일 16시 21분 16초
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금 10시 : 25분
분석화학 - Oral Presentation of Young Analytical Chemists
저자 및
성균관대학교 전자전기컴퓨터공학과, Korea
Phosphorylation by protein kinases is a predominant form of protein regulation. Abnormal phosphorylation of protein is linked to several medical states. Mutations in particular protein kinases give rise to a number of disorders and many naturally occurring toxins and the pathogens exert their effects by altering the phosphorylation states of intracellular proteins. Understanding of the specificity and regulation of particular protein kinase is crucial for the development of drugs to treat diabetes or cancers. Tremendous efforts have been devoted to quantify the catalytic activities of these enzymes. Radioactive labelling with [γ-32P]ATP with protein or peptide substrates are the most common method for measuring cellular kinase activities. However, the electrochemical method offers sensitive, cost-effective, and portable assays for kinase activity. Herein, we report an electrochemical assay for protein kinase A activity using a synthetic chemosensor. The synthetic chemosensor comprises of a bis(Zn2+-dipicolylamine) group and a thiol group at the ends of a hydrocarbon chain. The former strongly and selectively captures the phosphorylated enzyme product, forming a bridge between the two Zn2+ cations. The thiol group forms a self-assembled monolayer on the gold surface. In the presence of protein kinase, the substrate undergoes phosphorylation, which subsequently binds to the chemosensor, and then the ferrocene tag of the enzymatic product generates strong oxidation current under voltammetry. We performed differential pulse voltammetry to study the electrochemical signal obtained. Upon voltammetry, the mixture produced a well-defined oxidation peak corresponding to that of ferrocene and enabled a very low limit of detection. Our method was successfully applied even in the presence of human blood, and modified to enable single-use, chip-based electrochemical assay for kinase activity.