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Identification of iron dependent atpase in whole particulate suspension of the rat intestinal mucosa



Identification of iron dependent atpase in whole particulate suspension of the rat intestinal mucosa



Journal of Nippon Medical School 51(2): 240-252



Intestinal Fe absorption is recognized to be supported mainly by an active transport system, which requires energy consumption. Fe2+ or Fe3+ dependent ATPase activity may participate in intestinal Fe absorption, as well as in the case of Ca2+ absorption, in which Ca2+ dependent ATPase plays an important role in the process. An attempt was made to identify the presence of Fe2+ or Fe3+ dependent ATPase in the rat intestinal mucosa. Whole particulate suspension (WPS) from rat intestinal mucosa was used as the enzyme source. Eight mM ascorbic acid (Asc) or 1 mM citric acid (Cit) was added to the incubation mixture so as to prevent spontaneous oxidation of Fe2+ or hydroxylation of Fe3+, respectively. The increase in ATPase activities were apparently observed as a function of Fe2+ concentration. The increase in Fe2+ dependent ATPase activities was higher in the presence of Asc than in conditions without Asc. Km values of Fe2+ ATPase activity were 0.4 mM or 5.3 mM in the case of Asc (+) or Asc (-), respectively, and there was a statistical difference between these values (P < 0.001). However, Fe3+ dependent ATPase activity was not observed in the presence of Cit in this system. The activities of Fe2+ dependent ATPase in the presence of Asc was .apprx. 1/2 of Mg2+- or Ca2+-ATPase activities in WPS. The dependency on metal ions was dominant for Fe2+ compared to Ca2+ or Mg2+. Fe2+ dependent ATPase activity was not sensitive to ouabain, theophylline and L-phenylalanine at concentrations showing inhibitory effects. 32P-phosphorylated protein was identified from WPS in the presence of [.gamma.-32P] ATP and Fe2+, and the MW of the protein was estimated to be 97,000 by SDS [sodium dodecyl sulfate] polyacrylamide gel electrophoresis. The presence of Fe2+ dependent ATPase in the intestinal mucosa is suggested.

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