Functional roles of glutamic acid E143 and E705 residues in the N-terminus and transmembrane domain 7 of Anoctamin 1 in calcium and noxious heat sensing

Abstract

Anoctamin 1 (ANO1) is an anion channel that is activated by changes in cytosolic Ca²⁺ concentration and noxious heat Although the critical roles of ANO1 have been elucidated in various cell types, the control of its gating mechanisms by Ca²⁺ and heat remain more elusive. To investigate critical amino acid residues for modulation of Ca²⁺ and heat sensing, we constructed a randomized mutant library for ANO1. Among 695 random mutants, reduced Ca²⁺ sensitivity was observed in two mutants (mutant 84 and 87). Consequently, the E143A mutant showed reduced sensitivity to Ca²⁺ but not to high temperatures, whereas the E705V mutant exhibited reduced sensitivity to both Ca²⁺ and noxious heat These results suggest that the glutamic acids (E) at 143 and 705 residues in ANO1 are critical for modulation of Ca²⁺ and/or heat responses. Furthermore, these findings help to provide a better understanding of the Ca²⁺-mediated activation and heat-sensing mechanism of ANO1.