Halothane, Isoflurane, and Sevoflurane Increase the Kinetics of Ca2+-Induced Conformational Change of Recombinant Human Cardiac Troponin C

BACKGROUND: Halothane, isoflurane, and sevoflurane exert negative inotropic side effects, generally mediated via a reduced availability of intracellular calcium. Other possible mechanisms include modified intracellular calcium handling, impaired actomyosin cross-bridge cycling, and/or alteration of calcium-induced conformational changes of the regulatory troponin complex. METHODS: We investigated the effect of halothane, isoflurane, and sevoflurane on calcium-dependent kinetics of isolated human recombinant cardiac troponin C labeled with IAANS (HrcTnC(IAANS)) using stopped-flow and calcium titration techniques. RESULTS: Calcium concentration at half-maximal fluorescence intensity (K(d)) in the control group was 2.1 +/- 0.1 mM. Volatile anesthetics increased calcium sensitivity in a concentration-dependent fashion sevoflurane (K(d) 1.5-1.7 mM, P = 0.001) > halothane (K(d) 1.7-1.9 mM, P < 0.01) > isoflurane (K(d) 1.8-1.9 mM, P < 0.05). The rate constant of conformational changes after rapid dissociation of calcium from HrcTnC(IAANS) (k(off(c))) was moderately prolonged at 4 degrees C by halothane and isoflurane > sevoflurane. CONCLUSION: These mechanisms may counteract the effects of lower calcium availability, and can be responsible for abbreviated, and possibly incomplete, relaxation of cardiac muscle fibers in the presence of volatile anesthetics.