Mallikaratchy Prabodhika, Stahelin Robert V, Cao Zehui, Cho Wonhwa, Tan Weihong
Center for Research at the Bio/nano Interface, Department of Chemistry, Shands Cancer Centre, University of Florida, Gainesville, FL 32611, USA.
Chem Commun (Camb). 2006 Aug 14(30):3229-31. doi: 10.1039/b604778e. Epub 2006 Jun 23.
Protein kinase Cs are a family of serine and threonine kinases that mediate a wide variety of cellular signalling processes such as cell growth, differentiation, apoptosis and tumor development. We have selected high-affinity DNA aptamers for PKCdelta by capillary electrophoresis based SELEX (systematic evolution of ligands by exponential enrichment, CE-SELEX). We have demonstrated that fluorescently tagged PB9 aptamer can specifically recognize PKCdelta under in vitro conditions. The Kd of the aptamer-protein binding is 122 nM. These aptamers will enable us to apply fluorescently labelled probes to study the spatiotemporal dynamics and activation of individual endogenous PKC isoforms during various cell signalling processes.
蛋白激酶C是丝氨酸和苏氨酸激酶家族，介导多种细胞信号传导过程，如细胞生长、分化、凋亡和肿瘤发展。我们通过基于毛细管电泳的SELEX（指数富集配体系统进化，CE-SELEX）为PKCδ筛选了高亲和力DNA适配体。我们已经证明，荧光标记的PB9适配体在体外条件下可以特异性识别PKCδ。适配体与蛋白质结合的解离常数为122 nM。这些适配体将使我们能够应用荧光标记探针来研究各种细胞信号传导过程中单个内源性PKC亚型的时空动态和激活情况。
