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Intraluminal calcium dependence of calcium and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum calcium release


Intraluminal calcium dependence of calcium and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum calcium release



Journal of Biological Chemistry 267(29): 20850-20857



ISSN/ISBN: 0021-9258

The action of ryanodine upon sarcoplasmic reticulum (SR) Ca-2+ handling is controversial with evidence for both activation and inhibition of SR Ca-2+ release. In this study, the role of the intraluminal SR Ca-2+ load was probed as a potential regulator of ryanodine-mediated effects upon SR Ca-2+ release. Through dual-wavelength spectroscopy of Ca-2+:antipyrylazo III difference absorbance, the intraluminal Ca-2+ dependence of ryanodine and Ca-2+-induced Ca-2+ release (CICR) from skeletal SR vesicles was examined. Ryanodine addition after initiation of Ca-2+ uptake (a) increased the intraluminal Ca-2+ sensitivity of CICR and (b) stimulated spontaneous Ca-2+ release with a delayed onset. These ryanodine effects were inversely proportional to the intraluminal Ca-2+ load. Ryanodine also inhibited subsequent CICR after reaccumulation of Ca-2+ released from the initial CICR. These results provide evidence that ryanodine inhibits transitions between low and high affinity Ca-2+ binding states of an intraluminal Ca-2+ compartment, possibly calsequestrin. Conformational transitions of calsequestrin may be reciprocally coupled to transitions between open and closed states of the Ca-2+ release channel.

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