[Demonstration and properties of a Cl-/HCO3--ATPase in the avian salt gland (author's transl)].
An anion-stimulated, Mg2+-dependent, ouabain-insensitive ATPase is present in salt gland homogenates of domestic ducks (Anas platyrhynchos). The enzyme is unspecifically stimulated by various inorganic and organic anions including amino and sulfonic acids which are often used as buffer components (e. g. histidine, Bicin, PIPES, MES and HEPES). Therefore, the demonstration of ATPase stimulation by chloride strongly depends on the type and concentration of the buffer used and may also largely interfere with the stimulation caused by other anions present in the incubation medium. Of the inorganic anions tested chloride and bicarbonate appear to be the favorite physiological activators, but the possible role of carbonic acids in the stimulation of the anion-dependent ATPase should not be neglected. Km values are approximately 5.8 mM for Cl- and approximately 8.7 mM for HCO-3-activation. Maximal ATPase stimulation is obtained at 25 mM Cl- and approximately 30 mM HCO-3, respectively. The simultaneous presence of bicarbonate decreases chloride affinity and Vmax, and shifts the chloride optimum to lower concentrations. ATP is the most preferred substrate. Maximal activation by Cl- and HCO-3 occurs at ATP concentrations between 0.5 and 1 mM. ATP affinity increases in the presence of Cl- and HCO-3, respectively. Both chloride and bicarbonate require a Mg2+ to ATP ratio of approximately 0.5 and a pH value of 8.0 to 8.5 for optimal stimulation. Stimulation by Cl- and HCO-3 is inhibited by thiocyanate, cyanate and by the diuretic drugs furosemide, ethacrynic acid and mersalyl. Incubation media adapted for the simultaneous demonstration of both chloride and bicarbonate activation contained 10 to 20 mM histidine-Tris buffer at pH 8.0 to 8.5, 150 mM sucrose, 0.2 mM ouabain, 0.5 mM magnesium acetate, 1 mM ATP, (pH adjusted to 8.0-8.5 with Tris or NaOH), with and without 25 mM sodium chloride or 25 mM sodium bicarbonate.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D000838:Anions / Q000494:pharmacology; D001639:Bicarbonates / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism; D004372:Ducks; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D012491:Salt Gland / Q000201:enzymology*; D013379:Substrate Specificity
D000838:Anions; D001639:Bicarbonates; D002413:Cations, Divalent; D002712:Chlorides; D000251:Adenosine Triphosphatases; D008274:Magnesium
Nachweis und Eigenshaften einer Cl-/HCO3--ATPase in der Vogelsalzdrüse.