Characterizing the Dynamic Regulation of the K2P ion channel, TRESK

Summer 2017

Ian Tayler : MCB Neurobiology

Donor: Pergo Fund
Mentor: Stephen Brohawn

An essential component to better understanding cellular electrical signaling is to address how the resting membrane potential in neurons is established and modified to affect excitability. Two-pore domain potassium (K2P) ion channels are directly involved in this process and comprise a unique protein family that is essential for the maintenance of this resting membrane potential.
The TWIK-related spinal cord K+ channel (TRESK) is considered a major contributor to background K+ currents and is expressed abundantly in DRG neurons. This K2P channel is thought to be involved in pain sensation and sensory transduction, where its down-regulation may induce pain disorders such as allodynia and neuropathy, and loss of TRESK channel activity is strongly correlated to hereditary migraines with aura in humans. The goal of my research is to characterize the molecular structure of TRESK to understand the physical mechanism by which this channel operates.
This characterization will begin with the expression and purification of TRESK genes from a multitude of species that will be screened via fluorescence size-exclusion chromatography (FSEC). Promising constructs will be scaled up and purified using consecutive metal affinity and size-exclusion chromatography to isolate the TRESK channel from all other molecules in our expression systems. Purified channels will then be reconstituted into artificial bilayers, a reductionist system that allows us to study how TRESK activity changes in response to particular regulatory elements involved in its associated signaling pathways. This is an essential step to characterize the basis of the channel’s function and to ensure the integrity of our purified components before embarking on structural studies using X-ray crystallography, currently the most powerful technique for determining atomic resolution structures of biological macromolecules.

I would like to sincerely thank the Pergo Foundation for providing me with the financial support to conduct my research throughout the summer. Through this generous support, I have been allowed the opportunity to explore my passion for biological research in an incredibly immersive and comprehensive learning environment. This summer represented a profound period of personal growth in my life; being able to dive head first into a particular realm of academic knowledge was initially a daunting endeavor, and I questioned whether I could uphold my own personal expectations, as well as the expectations that my faculty mentor had of me. Thankfully, sheer excitement easily superseded my initial feelings of self doubt, and I quickly began to realize that I was in love with the process of academic research. During the coming months, I spent countless hours pouring through scientific literature as a means to learn about ways that I could bolster my own research in lab. In doing so, I began to realize just how fulfilling it was to formulate a question, and then seek various methods of conceptualization and implementation to to answer the question. This experience vastly differed from the conventional process of learning in a class environment, where textbooks and other course materials are simply used to supplement information presented in class. I found that delving into scientific papers to glean information about cutting-edge topics was enriching to my mind, spirit, and, ultimately, led to a greater understanding of big-picture concepts and ideas that may one day perfuse from my own research. I am so immensely grateful for the opportunity to have learned what it means to be part of an intellectual community engaged in research. To that end, I would like to extend my deepest appreciation to those at the Pergo Foundation for enabling such an enriching and transformative experience.