Optimizing Transfection, Expression, and Purification of the Human Transient Receptor Potential Melastatin 8 (hTRPM8) from HEK293 cells

Transient Receptor Potential Melastatin 8 (TRPM8) is a non-selective cation channel notable as a primary cold sensor in humans. It is also involved in a variety of (patho)physiological events including pain sensation, chronic cough, diabetes, obesity, and cancer. TRPM8 is modulated by a variety of stimuli including pH, temperature, cooling agents, voltage, lipid, and other proteins. However, the molecular mechanism underlining its function has not yet clear raising the need for isolated proteins to be well-characterized. Over 20 years, E. coli has been a heterologous expression system of interest due to its low cost and high yield. However, the lack of post-translational modifications and chaperone may cause a misfolding or affect protein function. Mammalian expression system addresses these drawbacks and is a good candidate for the functional study of complex human protein. Here I describe my research in optimizing the transfection, expression, and purification of the human TRPM8 from adherent Human Embryonic Kidney (HEK293) cells which can be used for small-scale studies including, but not limited to, planar lipid bilayer electrophysiology.