Structural Study of Human Voltage-Gated Potassium Channel Kv1.1

Voltage-gated ion channels are vital components to the process of signal transmission in nerve cells. They constitute an inviting target for the development of drugs which target aberrant pain signal transmission. Voltage-gated potassium channels are an essential component in signalling along neural pathways, as their role in restoring resting membrane potential in nerve cells is crucial to priming neurons for further signal transmission following membrane depolarisation. In this thesis, work towards developing a structural understanding of interactions between the human Kv1.1 voltage-gated potassium channel and inhibitory molecules will be discussed. This project has involved the development of expression vectors and cell lines for expression of Kv1.1 in human cell cultures, as well as experimentation with methods for extracting the protein with recently developed maleic-acid polymers SMA and DIBMA, which retain the native lipid envelop surrounding the channel complex. The extracted protein has been analysed through both biochemical characterisation though mass spectrometry and circular dichroism spectroscopy, as well as structural studies conducted through both negative stain and cryo-EM, resulting a low-resolution model from negative stain. This work lays the foundation for future investigation of the human Kv1.1 channel, providing tested means and methodologies for the production of proteins in human cell expression systems, extraction through non-detergent methods and their analysis via cryo-electron microscopy.