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To unequivocally determine the residue-specific consequences of acetylation on Tn-Tpm-based legislation of actomyosin task, we evaluated the effects of K326Q and K328Q acetyl (Ac)-mimetic actin on Ca2+-dependent, in vitro motility variables of reconstituted thin filaments (RTFs). Incorporation of K328Q actin significantly enhanced Ca2+ sensitivity of RTF activation in accordance with control. Together, our results claim that actin acetylation, specially Lys328, modul