Abstract:Here, we report how the mutagenesis of the -bulge residues (D137 and W138) in mNeonGreen, a bright fluorescent protein, unlocks and tunes the anion preference at physiological pH for sulfate, resulting in the turn-off sensor SulfOFF-1. This unprecedented sensing arises from an enhancement in the kinetics of binding, largely driven by position 138. In line with these data, molecular dynamics (MD) simulations capture how the coordinated entry and gating of sulfate into the -barrel is eliminated upon mutagenesi… Show more
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