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LRET measurements were performed using sensitized emission lifetime measurements from receptors tagged with terbium chelate and ATTO 465, measured using double cysteine mutants at residues 139 and 340 in HEK293T cells (a) and oocyte membranes (b) at pH 8 (black) and pH 6 (red). The corresponding donor lifetime measurements are shown in c and d, respectively. Inset, positions of sites 139 and 340 in the extracellular domain of a single subunit of the ASIC.
