AUTHOR=Little Sean C., Tikunova Svetlana B., Swartz Darl R., Norman Catalina , Davis Jonathan P. TITLE=Measurement of Calcium Dissociation Rates from Troponin C in Rigor Skeletal Myofibrils JOURNAL=Frontiers in Physiology VOLUME=2 YEAR=2011 URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2011.00070 DOI=10.3389/fphys.2011.00070 ISSN=1664-042X ABSTRACT=

Ca2+ dissociation from the regulatory domain of troponin C may influence the rate of striated muscle relaxation. However, Ca²⁺ dissociation from troponin C has not been measured within the geometric and stoichiometric constraints of the muscle fiber. Here we report the rates of Ca²⁺ dissociation from the N-terminal regulatory and C-terminal structural domains of fluorescent troponin C constructs reconstituted into rabbit rigor psoas myofibrils using stopped-flow technology. Chicken skeletal troponin C fluorescently labeled at Cys 101, troponin C^IAEDANS, reported Ca²⁺ dissociation exclusively from the structural domain of troponin C at ∼0.37, 0.06, and 0.07/s in isolation, in the presence of troponin I and in myofibrils at 15°C, respectively. Ca²⁺ dissociation from the regulatory domain was observed utilizing fluorescently labeled troponin C containing the T54C and C101S mutations. Troponin CMIANST54C,C101S reported Ca²⁺ dissociation exclusively from the regulatory domain of troponin C at >1000, 8.8, and 15/s in isolation, in the presence of troponin I and in myofibrils at 15°C, respectively. Interestingly, troponin CIAANST54C,C101S reported a biphasic fluorescence change upon Ca²⁺ dissociation from the N- and C-terminal domains of troponin C with rates that were similar to those reported by troponin CMIANST54C,C101S and troponin C^IAEDANS at all levels of the troponin C systems. Furthermore, the rate of Ca²⁺ dissociation from troponin C in the myofibrils was similar to the rate of Ca²⁺ dissociation measured from the troponin C-troponin I complexes. Since the rate of Ca²⁺ dissociation from the regulatory domain of TnC in myofibrils is similar to the rate of skeletal muscle relaxation, Ca²⁺ dissociation from troponin C may influence relaxation.