Pannexin channels are a family of recently discovered membrane proteins found in nearly every tissue of the human body. These channels have been classified as large ‘pore forming’ proteins which, when activated, create a passageway through the cell membrane through which ions and molecules transit. Current literature suggests that the actual pannexin channel is formed from a hexameric arrangement of individual monomeric pannexin subunits, resulting in a central permeation pathway for conducting ions. Opening of pannexin channels can be accomplished through several mechanisms. During apoptosis, for example, cleavage of the pannexin C-terminal domain results in a constitutively open channel through which ATP is released. However, curiously, pannexins have also been known to be activated by a variety of other stimuli such as cellular depolarization, exposure to signaling ions like Ca2+ and K+, and interacting with various other membrane receptors like members of the ATP-sensing P2X and P2Y family. How can pannexin channels sense and respond to such a diverse array of stimuli, and what is the fundamental ‘gating process’ that defines channel opening? Here, we use electrophysiology to study the activation of pannexin-1 (Panx1). We used a protein chimera approach to identify that the first extracellular domain of Panx1 is critical for inhibitor action. Mutagenesis of this region identified that bulky hydrophobic amino acids in this region confer sensitivity of the channel to various drug compounds. We also identified that the very N-terminus of Panx1 is important for voltage sensing, and that subtle modifications of the N-terminus results in channels with altered channel gating when exposed to voltage stimulation. Finally, we made attempts to solve the structure of a Panx1 channel. Iterative rounds of optimization yielded crystals that diffract x-rays just beyond 5 Å.