Different degrees of cooperativity of the Ca2+-induced changes in fluorescence intensity of solubilized sarcoplasmic reticulum ATPase.

TL;DRAbstract

The tryptophan fluorescence emission of sarcoplasmic reticulum Ca2+-ATPase was studied both in purified ATPase vesicles and in ATPase solubilized with the nonionic detergent dodecyloctaethyleneglycolmonoether (C12E8). Fluorescence intensity changes in purified ATPase were titrated as a function of free Ca2+ in the medium. It exhibited a cooperative pattern, with a Hill number of 2.21 +/- 0.02 and K0.5 = 0.51 microM Ca2+. Upon solubilization of the ATPase, the cooperative pattern of fluorescence change was lost; the Hill number was 0.96 and K0.5 = 1.4 microM Ca2+. When solubilization was carried out in the presence of 0.5 or 1.0 mM CaCl2, followed by the titrations of fluorescence change in the micromolar Ca2+ range, the cooperative pattern was preserved under the same concentrations of C12E8 which would otherwise promote the loss in cooperativity. For the ATPase solubilized in millimolar Ca2+, the Hill number was 1.98 with a K0.5 = 1.5 microM Ca2+. The maximal amount of Ca2+ bound to t

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