Calcium uptake by isolated sarcoplasmic reticulum treated with dithiothreitol.

Isolated vesicles of the sarcoplasmic reticulum are known to take up calcium when provided with magnesium adenosine triphosphate as an energy source. Preparations of high activity are obtained by keeping the vesicles in 5 millimolar dithiothreitol (a reagent that reduces disulfide groups), and these preparations retain activity for a week or longer. The highly active preparations lend themselves to a spectrophotometric method for following calcium uptake, and continuous uptake kinetics are readily obtained. Calcium uptake appears to follow Michaelis-Menten kinetics (K(m) = 8x10(-6); V(mas), = 7x10(-7) mole per second per milligram of protein). These preparations are also useful for studying the effects of inhibitors of uptake, such as quinine. When extrapolated to the intact muscle, the results from these isolated vesicles should give a better estimate than has been available of the actual rates of calcium uptake and of the physiological effect of inhibitors of uptake.