Preparation of the Transient Receptor Potential Vanilloid 2 (TRPV2) channel for structural studies

Transient Receptor Potential (TRP) channels play numerous important physiological roles in humans. Notably, they are involved in temperature sensing and regulation, in the proper functioning of immune and cardiac systems, in skin, hair, and bone physiology and in many types of cancer. Because of their physiological significance there has been much interest in elucidating their molecular mechanisms of action. Recent improvements in eukaryotic protein expression and purification techniques and in cryo-electron microscopy (cryo-EM) have greatly facilitated TRP channel studies. The TRP Vanilloid 2 (TRPV2) channel has emerged as particularly amenable to structural studies and its structure has been solved by both X-ray crystallography and by cryo-EM. Here, we provide an overview of demands posed by X-ray crystallography and cryo-EM on protein sample preparation and outline a step-by-step protocol for preparing the TRPV2 protein for structure determination by both of these techniques.