OPTIMIZATION OF EXPRESSION AND PURIFICATION OF THE HUMAN TRPM8 ION CHANNEL

The human transient receptor potential melastatin 8 (TRPM8) ion channel plays important and diverse roles in human physiology, including as the primary human cold sensor. Additionally, TRPM8 has also been implicated in a variety of human diseases including cancer, diabetes, and obesity. TRPM8 is modulated by a variety of stimuli, including temperature, pH, voltage, lipids, chemicals, and other proteins. From its discovery in 2001, interest has emerged in better understanding the molecular underpinnings of TRPM8 function given its physiological roles and potential for therapeutic intervention. Post-translational modifications have been implicated in some aspects of TRPM8 function. To better understand the role of post-translational modifications in function, the human TRPM8 protein is expressed and purified from mammalian cells (HEK293) and from bacterial cells (E. coli) which allow for comparative experiments to better decipher the role of post-translational modifications on TRPM8 function. The mammalian and bacterial expressed human protein with and without posttranslational modifications is subjected to planar bilayer electrophysiology experiments. The results of these studies will be discussed in addition to the process of optimizing the bacterial and mammalian expression and purification of this relatively large human membrane protein.