Relating the binding of single ligands to activation in single HCN and CNG channels (A05)

单标记cGMP和cAMP分子在CNGA2和HCN2离子通道中的相互作用将被研究。为了在高纳摩尔浓度下实现这一目标，通道将在放置在消失场中的支持天然膜或位于亚衍射尺寸的膜片移液器尖端的切除膜片中进行研究，以平行记录单通道电流。为了优化这些方法，我们将在硅上设计、合成和表征新的cGMP和cAMP衍生物。观察结果将根据隐马尔可夫模型进行分析，以完善我们对渠道动力学的理解。

The interactions of single labeled cGMP and cAMP molecules will be studied in CNGA2 and HCN2 ion channels. To achieve this at high nanomolar concentrations, the channels will be studied in either supported native membranes placed in an evanescent field or in excised membrane patches positioned at the tip of patch pipettes with sub-diffraction dimensions to record single-channel currents in parallel. To optimize these methods novel cGMP and cAMP derivatives will be designed in silico, synthesized and characterized. The observations will be analyzed in terms of hidden Markov models to refine our understanding of the channel dynamics.