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. Author manuscript; available in PMC: 2013 Aug 14.

Published in final edited form as: Biochemistry. 2012 Aug 1;51(32):6413–6420. doi: 10.1021/bi3006835

Fig. 1 — Effects of Tm-FHC mutants on actin-S1 ATPase activity. Data obtained by continuously monitoring Pi liberation (see Methods). A. Effects of Tm and Tn +/− Ca²⁺. ATPase was monitored during successive additions of S1, actin, Tm, Tn, EGTA to a solution containing, 0.1mM CaCl₂, 15 mM NaCl, 3 mM MgCl₂, 1 mM DTT and 1 mM ATP in buffer (10 mM HEPES, pH 7.5). Data were normalized to actin-S1 activity after subtracting the background S1 ATPase. B. ATPase vs. [Ca²⁺]. CaCl₂ was titrated into the reconstituted actinTmTn thin filament in buffer containing 1mM

EGTA and 1 mM NTA. Data were normalized to the high [Ca²⁺] values fitted to the Hill equation. Background S1 alone ATPase was subtracted.

Fig. 1 — Effects of Tm-FHC mutants on actin-S1 ATPase activity. Data obtained by continuously monitoring Pi liberation (see Methods). A. Effects of Tm and Tn +/− Ca²⁺. ATPase was monitored during successive additions of S1, actin, Tm, Tn, EGTA to a solution containing, 0.1mM CaCl₂, 15 mM NaCl, 3 mM MgCl₂, 1 mM DTT and 1 mM ATP in buffer (10 mM HEPES, pH 7.5). Data were normalized to actin-S1 activity after subtracting the background S1 ATPase. B. ATPase vs. [Ca²⁺]. CaCl₂ was titrated into the reconstituted actinTmTn thin filament in buffer containing 1mM

EGTA and 1 mM NTA. Data were normalized to the high [Ca²⁺] values fitted to the Hill equation. Background S1 alone ATPase was subtracted.