C-terminal Acidic Cluster Is Involved in Ca2+-induced Regulation of Human Transient Receptor Potential Ankyrin 1 Channel

The transient receptor potential ankyrin 1 (TRPA1) channel is a Ca2+-permeable cation channel whose activation results from a complex synergy between distinct activation sites, one of which is especially important for determining its sensitivity to chemical, voltage and cold stimuli. From the cytoplasmic side, TRPA1 is critically regulated by Ca2+ ions, and this mechanism represents a self-modulating feedback loop that first augments and then inhibits the initial activation. We investigated the contribution of the cluster of acidic residues in the distal C terminus of TRPA1 in these processes using mutagenesis, whole cell electrophysiology, and molecular dynamics simulations and found that the neutralization of four conserved residues, namely Glu1077 and Asp1080–Asp1082 in human TRPA1, had strong effects on the Ca2+- and voltage-dependent potentiation and/or inactivation of agonist-induced responses. The surprising finding was that truncation of the C terminus by only 20 residues selectively slowed down the Ca2+-dependent inactivation 2.9-fold without affecting other functional parameters. Our findings identify the conserved acidic motif in the C terminus that is actively involved in TRPA1 regulation by Ca2+. TRPA1 channel is modulated by Ca2+, but the molecular mechanisms are unclear. Mutations in the distal C-terminal acidic domain altered Ca2+ dependence of TRPA1. The C-terminal acidic cluster is involved in the Ca2+-induced potentiation and inactivation of TRPA1. Identification of the Ca2+-dependent domain is important for understanding the role of TRPA1 in chemical nociception.