Part I (Chapters 1-4)Pannexins are a family of proteins that allows for ATP release through its central pore. The ATP released through this mechanism allows for many downstream signaling events. Pannexins have been shown to play pathophysiological roles in events such as stroke, epileptic seizure, and neuropathic pain, making them excellent targets for drug discovery and clinical studies. Despite this, the methods of activation, deactivation, and inhibition of these channels remains poorly understood. This can largely be attributed to the lack of structural and mechanistic characterization of these channels. In this work, purification of multiple Pannexin subtypes and constructs is established and significant work towards obtaining structure models is completed. Further structural analysis also gives insight into how various pharmacological reagents may be inhibiting these channels. This work makes significant steps towards developing a platform to understand the molecular mechanisms governing this biomedically relevant family of proteins. Part II (Chapters 5-8) Tweety Homologues (TTYH1-3) comprise a novel family of proteins with roles in critical biological processes such as neurogenesis and embryonic development. TTYHs have been shown to play pathophysiological roles in glioma formation, renal cancer, and gastric cancer. This growing body of research highlights TTYH’s promising role as a therapeutic target. Despite TTYH’s obvious clinical relevance, research into these protein’s therapeutic potential is largely uninvestigated. This can be largely attributed to the lack of understanding regarding TTYHs molecular role. In this work, purification of the TTYH family is established and significant work towards obtaining a structural model is completed. The isolated protein will provide a powerful tool for probing TTYHs functional role in vitro. The work towards obtaining structural models will prove useful for determination of molecular regulation mechanisms that are essential for therapeutic studies.