Extracellular Subunit Interactions Control Transitions Between Functional States of Acid-sensing Ion Channel 1a [Membrane Biology]

June 12th, 2015 by Gwiazda, K., Bonifacio, G., Vullo, S., Kellenberger, S.

Acid-sensing ion channels (ASICs) are neuronal, voltage-independent Na+ channels that are transiently activated by extracellular acidification. They are involved in pain sensation, the expression of fear and in neurodegeneration after ischemic stroke. Our study investigates the role of extracellular subunit interactions in ASIC1a function. We identified two regions involved in critical intersubunit interactions. First, formation of an engineered disulfide bond between the palm and thumb domains leads to partial channel closure. Second, linking Glu235 of a finger loop to either one of two different residues of the knuckle of a neighboring subunit opens the channel at physiological pH or disrupts its activity. This suggests that one finger-knuckle disulfide bond (E235C/K393C) sets the channel in an open state, while the other (E235C/Y389C) switches the channel to a non-conducting state. Voltage-clamp fluorometry experiments indicate that both the finger loop and the knuckle move away from the β-ball residue Trp233 during acidification and subsequent desensitization. Together, these observations reveal that ASIC1a opening is accompanied by a distance increase between adjacent thumb and palm domains, as well as a movement of Glu235 relative to the knuckle helix. Our study identifies subunit interactions in the extracellular loop and shows that dynamic changes of these interactions are critical for normal ASIC function.