The effect of temperature on some calcium-binding properties of troponin C and calmodulin.
Some calcium-binding properties of skeletal and cardiac troponin C (TnC) have been measured as functions of temperature employing several physical and spectroscopic techniques. The degree of exposure of the tyrosine residues in brain calmodulin has also been determined by a new approach. Circular dichroism thermal unfolding profiles have been established for the three cases: metal-free protein, high-affinity sites filled, and fully saturated. In addition some thermodynamic parameters have been calculated for these reversible melting process. It was found that the calcium-binding parameters n and K, where n is the fraction of the total conformational change and K is the apparent association constant, for both skeletal and cardiac TnC, did not vary significantly over the temperature range 10-38 degrees C, but at 50 degrees C differences became quite apparent, dramatically so in the case of the skeletal protein. The technique of thermal perturbation difference spectroscopy was applied to determine the degree of exposure of aromatic chromophores for the TnC(s) and calmodulin in the absence and presence of calcium. For skeletal TnC and calmodulin the results were in good agreement with previous observations, but the reduced degree of exposure of the tyrosine residues in cardiac TnC, in the absence of Ca2+, was contrary to the earlier work. Calcium-induced difference absorption spectra have been measured for the TnC(s) over the temperature range 10-70 degrees C. Cardiac TnC showed greater heat stability than its skeletal counterpart, in terms of the rate and the amount of change of the difference spectral maxima.
D000818:Animals; D001842:Bone and Bones / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D009124:Muscle Proteins / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D011489:Protein Denaturation; D011817:Rabbits; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013816:Thermodynamics; D014336:Troponin / Q000378:metabolism*; D019209:Troponin C
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D009124:Muscle Proteins; D014336:Troponin; D019209:Troponin C; D002118:Calcium