ANALYSIS OF PROTEIN KINASE A A KINASE ANCHORING PROTEIN INTERACTIONS

越来越多的证据表明联合本地化的重要性 激酶及其底物被细胞吸附以获得特定的生物 效果。这种共址似乎是通过脚手架或 锚定蛋白共同定位适当的激酶和 底物。A激酶锚定蛋白(AKAP)中的一种这样的蛋白质 它共定位蛋白激酶A(PKA)及其底物 神经生理素。这个项目的目标是开发一个流程 细胞计数法检测AKP-PKA相互作用。生物素 标记的AKAP已经生产出来，并将被附着到微球上。 PKA调节亚基将被荧光标记并与其结合 用流式细胞仪检测固定化AKAP。

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.