Production of K2P2.1 (TREK-1) for structural studies

Abstract

K_2P (KCNK) potassium channels form 'background' or 'leak' currents that are important for controlling cell excitability in the brain, cardiovascular system, and somatosensory neurons. K_2P2.1 (TREK-1) is one of the founding members of this family and one of the first well-characterized polymodal ion channels capable of responding to a variety of physical and chemical gating cues. Of the six K_2P subfamilies, the thermo-and mechano-sensitive TREK subfamily comprising K_2P2.1 (TREK-1), K_2P4.1 (TRAAK), and K_2P10.1 (TREK-2) is the first to have structures determined for each subfamily member. These structural studies have revealed key architectural features that provide a framework for understanding how gating cues sensed by different channel elements converge on the K_2P selectivity filter C-type gate. TREK family structural studies have also revealed numerous sites where small molecules or lipids bind and affect channel function. This rich structural landscape provides the framework for probing K_2P function and for the development of new K_2P-directed agents. Such molecules may be useful for affecting processes where TREK channels are important such as anesthesia, pain, arrythmia, ischemia, migraine, intraocular pressure, and lung injury. Production of high quality protein samples is key to addressing new questions about K_2P function and pharmacology. Here, we present methods for producing pure K_2P2.1 (TREK-1) suitable for advancing towards these goals through structural and biochemical studies.

Keywords: C-type gate; K(2P) channel; Selectivity filter; X-ray crystallography.