ANALYSIS OF PROTEIN KINASE A A KINASE ANCHORING PROTEIN INTERACTIONS

越来越多的证据表明， 激酶及其底物通过细胞获得特异性生物活性 方面的影响. 这种共定位似乎是由支架或 锚定蛋白，其共定位适当的激酶， 印刷受体. A激酶介导蛋白（AKAP） 其共定位蛋白激酶A（PKA）及其底物 神经垂体素 这个项目的目标是开发一个流程 用于测量AKP-PKA相互作用的细胞计数测定。 生物素 标记的AKAP已经产生并将附着到微球上。 PKA调节亚基将被荧光标记，其结合 将通过流式细胞术测量固定化AKAP的浓度。

Growing evidence points to the importance of colocalization of kinases and their substrates by cells to acheive specific biological effects. Such colocalization appears to be mediated by scaffolding or anchoring proteins which colocalize the appropriate kinases and substrates. One such protein in the A Kinase Anchoring Protein (AKAP) which colocalizes protein kinase A (PKA) and its substrate neurophysin. The object of this project is to develop a flow cytometric assay for the measurement of AKP-PKA interactions. Biotin tagged AKAP has been produced and will be attached to microspheres. PKA regulatory subunit will be fluorescently labeled and its binding to immobilized AKAP will be measured by flow cytometry.