Ca-activated ATPase activity in subcellular fractions of mouse pancreatic islets.
Ca-stimulated ATPase activity has been demonstrated in homogenates of mouse pancreatic islets. On subcellular fractionation Ca-ATPase activity was found in secretory granules, mitochondria, and microsomes, but not in the postmicrosomal fractions. Highest specific activity was found in the granules. In all active subcellular fractions two Km(Ca) values for Ca-ATPase around 7.0 X 10(-6) and 1.8 X 10(-7) M were estimated. Assuming an ATP hydrolysis:Ca pumping ratio of 1:2, the highest capacity for active Ca transport was found in secretory granules and mitochondria. Concentrations of 40 mM or higher of Na and 10(-5) M cyclic AMP inhibited Ca-ATPase in all subfractions. Caffeine at a concentration of 10 mM inhibited Ca-ATPase significantly in secretory granules and microsomes. Also MG-ATPase activity was demonstrated in the various subfractions. This activity was compared with that of Ca-ATPase at identical concentrations of free metal ions and in the absence or presence of various inhibitors. It was concluded that high-affinity Ca-ATPase and Mg-ATPase are two different enzymic entities. Ca-ATPase may tentatively be assumed to participate in active transport of Ca between intracellular compartments and to constitute a Ca-accumulating system which returns the cytosolic free Ca concentration to the resting state after stimulation of the beta-cells by secretagogues. This enzyme may therefore play a significant role in regulation of insulin release.
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000628:Aminophylline / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D001693:Biological Transport, Active; D002110:Caffeine / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000201:enzymology; D007515:Islets of Langerhans / Q000201:enzymology* / Q000648:ultrastructure; D007700:Kinetics; D008297:Male; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D012964:Sodium / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D014970:Xanthines / Q000494:pharmacology
D014970:Xanthines; D000628:Aminophylline; D002110:Caffeine; D012965:Sodium Chloride; D002731:4-Chloromercuribenzenesulfonate; D012964:Sodium; D000242:Cyclic AMP