Effects of Mg 2+ on calcium accumulation by two fractions of sarcoplasmic reticulum from rabbit skeletal muscle

Abstract

Calcium accumulation by two fractions of sarcoplasmic reticulum presumably derived from longitudinal tubules (light vesicles) and terminal cisternae (heavy vesicles) was examined radiochemically in the presence of various free Mg 2+ concentrations. Both fractions of sarcoplasmic reticulum exhibited a Mg 2+-dependent increase in phosphate-supported calcium uptake velocity, though half-maximal velocity in heavy vesicles occurred at a much higher free Mg 2+ concentration than that in light vesicles (i.e., approx. 0.90 mM vs. approx. 0.02 mM Mg 2+). Calcium uptake velocity in light vesicles correlated with Ca 2+-dependent ATPase activity, suggesting that Mg 2+ stimulated the calcium pump. Calcium uptake velocity in heavy vesicles did not correlate with Ca 2+-dependent ATPase activity, although a Mg 2+-dependent increase in calcium influx was observed. Thus, Mg 2+ may increase the coupling of ATP hydrolysis to calcium transport in heavy vesicles. Analyses of calcium sequestration (in the absence of phosphate) showed a similar trend in that elevation of Mg 2+ from 0.07 to 5 mM stimulated calcium sequestration in heavy vesicles much more than in light vesicles. This difference between the two fractions of sarcoplasmic reticulum was not explained by phosphoenzyme (EP) level or distribution. Analyses of calcium uptake, Ca 2+-dependent ATPase activity, and unidirectional calcium flux in the presence of approx. 0.4 mM Mg 2+ suggested that ruthenium red (0.5 μ M) can also increase the coupling of ATP hydrolysis to calcium transport in heavy vesicles, with no effect in light vesicles. These functional differences between light and heavy vesicles suggest that calcium transport in terminal cisternae is regulated differently from that in longitudinal tubules.